concentrated tris base Search Results


trizma base 2 amino 2 hydroxymethyl 1 3 propanediol  (Thermo Fisher)
96
Thermo Fisher trizma base 2 amino 2 hydroxymethyl 1 3 propanediol
Trizma Base 2 Amino 2 Hydroxymethyl 1 3 Propanediol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/trizma base 2 amino 2 hydroxymethyl 1 3 propanediol/product/Thermo Fisher
Average 96 stars, based on 1 article reviews
trizma base 2 amino 2 hydroxymethyl 1 3 propanediol - by Bioz Stars, 2026-04
96/100 stars
  Buy from Supplier
unmasking solution  (Vector Laboratories)
96
Vector Laboratories unmasking solution
Unmasking Solution, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/unmasking solution/product/Vector Laboratories
Average 96 stars, based on 1 article reviews
unmasking solution - by Bioz Stars, 2026-04
96/100 stars
  Buy from Supplier
novex nupage precast tris base  (Thermo Fisher)
99
Thermo Fisher novex nupage precast tris base
Novex Nupage Precast Tris Base, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/novex nupage precast tris base/product/Thermo Fisher
Average 99 stars, based on 1 article reviews
novex nupage precast tris base - by Bioz Stars, 2026-04
99/100 stars
  Buy from Supplier
trizma base  (Thermo Fisher)
99
Thermo Fisher trizma base
( a ) The secondary structure of the ladder ribozyme and a schematic showing its function. The ribozyme is shown in red, whilst the substrate strands are shown in black. ( b ) A representative 8% urea PAGE stained with SYBR Gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 19-72 to R3C RNA (total monomer concentration = 1 mM, 10.5 µM substrate, 10.5 µM ribozyme) after a 2 hr reaction at 30°C in 50 <t>mM</t> <t>Tris-HCl</t> pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 0.75:1 (Lys) 19-72 :RNA conditions are shown in blue and red, respectively ( c ) Variation in absorbance at 500 nm as a measure of coacervate formation upon addition of varying ratios of (Lys) 19-72 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( d ) Example fluorescence microscopy image of (Lys) 19-72: RNA condensates at a ratio of 0.75:1 (Lys) 19-72 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( e ) Kinetics of chain elongation in solution (blue, first-order model), and with 0.75:1 (Lys) 19-72 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . A total RNA monomer concentration of 1 mM was achieved by combining 9.5 µM substrate, 1 µM Cy5-tagged substrate and 10.5 µM ribozyme. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( f ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 0.75:1 (Lys) 19-72 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Equivalent data for condensates formed from the shorter (Lys) 5-24 peptide is shown in . Figure 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Figure 1—source data 2. Numerical turbidity data for . Figure 1—source data 3. Unprocessed and uncropped fluorescence microscope image for 0.75:1 (Lys) 19-72 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the accompanying spreadsheet.
Trizma Base, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/trizma base/product/Thermo Fisher
Average 99 stars, based on 1 article reviews
trizma base - by Bioz Stars, 2026-04
99/100 stars
  Buy from Supplier
buffer exchange into tbs  (Thermo Fisher)
99
Thermo Fisher buffer exchange into tbs
( a ) The secondary structure of the ladder ribozyme and a schematic showing its function. The ribozyme is shown in red, whilst the substrate strands are shown in black. ( b ) A representative 8% urea PAGE stained with SYBR Gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 19-72 to R3C RNA (total monomer concentration = 1 mM, 10.5 µM substrate, 10.5 µM ribozyme) after a 2 hr reaction at 30°C in 50 <t>mM</t> <t>Tris-HCl</t> pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 0.75:1 (Lys) 19-72 :RNA conditions are shown in blue and red, respectively ( c ) Variation in absorbance at 500 nm as a measure of coacervate formation upon addition of varying ratios of (Lys) 19-72 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( d ) Example fluorescence microscopy image of (Lys) 19-72: RNA condensates at a ratio of 0.75:1 (Lys) 19-72 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( e ) Kinetics of chain elongation in solution (blue, first-order model), and with 0.75:1 (Lys) 19-72 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . A total RNA monomer concentration of 1 mM was achieved by combining 9.5 µM substrate, 1 µM Cy5-tagged substrate and 10.5 µM ribozyme. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( f ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 0.75:1 (Lys) 19-72 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Equivalent data for condensates formed from the shorter (Lys) 5-24 peptide is shown in . Figure 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Figure 1—source data 2. Numerical turbidity data for . Figure 1—source data 3. Unprocessed and uncropped fluorescence microscope image for 0.75:1 (Lys) 19-72 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the accompanying spreadsheet.
Buffer Exchange Into Tbs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/buffer exchange into tbs/product/Thermo Fisher
Average 99 stars, based on 1 article reviews
buffer exchange into tbs - by Bioz Stars, 2026-04
99/100 stars
  Buy from Supplier
mri contrast agents  (Bracco Imaging Deutschland GmbH)
90
Bracco Imaging Deutschland GmbH mri contrast agents
( a ) The secondary structure of the ladder ribozyme and a schematic showing its function. The ribozyme is shown in red, whilst the substrate strands are shown in black. ( b ) A representative 8% urea PAGE stained with SYBR Gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 19-72 to R3C RNA (total monomer concentration = 1 mM, 10.5 µM substrate, 10.5 µM ribozyme) after a 2 hr reaction at 30°C in 50 <t>mM</t> <t>Tris-HCl</t> pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 0.75:1 (Lys) 19-72 :RNA conditions are shown in blue and red, respectively ( c ) Variation in absorbance at 500 nm as a measure of coacervate formation upon addition of varying ratios of (Lys) 19-72 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( d ) Example fluorescence microscopy image of (Lys) 19-72: RNA condensates at a ratio of 0.75:1 (Lys) 19-72 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( e ) Kinetics of chain elongation in solution (blue, first-order model), and with 0.75:1 (Lys) 19-72 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . A total RNA monomer concentration of 1 mM was achieved by combining 9.5 µM substrate, 1 µM Cy5-tagged substrate and 10.5 µM ribozyme. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( f ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 0.75:1 (Lys) 19-72 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Equivalent data for condensates formed from the shorter (Lys) 5-24 peptide is shown in . Figure 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Figure 1—source data 2. Numerical turbidity data for . Figure 1—source data 3. Unprocessed and uncropped fluorescence microscope image for 0.75:1 (Lys) 19-72 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the accompanying spreadsheet.
Mri Contrast Agents, supplied by Bracco Imaging Deutschland GmbH, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mri contrast agents/product/Bracco Imaging Deutschland GmbH
Average 90 stars, based on 1 article reviews
mri contrast agents - by Bioz Stars, 2026-04
90/100 stars
  Buy from Supplier
tris trizma base  (Fluka Chemical)
90
Fluka Chemical tris trizma base
( a ) The secondary structure of the ladder ribozyme and a schematic showing its function. The ribozyme is shown in red, whilst the substrate strands are shown in black. ( b ) A representative 8% urea PAGE stained with SYBR Gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 19-72 to R3C RNA (total monomer concentration = 1 mM, 10.5 µM substrate, 10.5 µM ribozyme) after a 2 hr reaction at 30°C in 50 <t>mM</t> <t>Tris-HCl</t> pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 0.75:1 (Lys) 19-72 :RNA conditions are shown in blue and red, respectively ( c ) Variation in absorbance at 500 nm as a measure of coacervate formation upon addition of varying ratios of (Lys) 19-72 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( d ) Example fluorescence microscopy image of (Lys) 19-72: RNA condensates at a ratio of 0.75:1 (Lys) 19-72 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( e ) Kinetics of chain elongation in solution (blue, first-order model), and with 0.75:1 (Lys) 19-72 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . A total RNA monomer concentration of 1 mM was achieved by combining 9.5 µM substrate, 1 µM Cy5-tagged substrate and 10.5 µM ribozyme. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( f ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 0.75:1 (Lys) 19-72 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Equivalent data for condensates formed from the shorter (Lys) 5-24 peptide is shown in . Figure 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Figure 1—source data 2. Numerical turbidity data for . Figure 1—source data 3. Unprocessed and uncropped fluorescence microscope image for 0.75:1 (Lys) 19-72 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the accompanying spreadsheet.
Tris Trizma Base, supplied by Fluka Chemical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/tris trizma base/product/Fluka Chemical
Average 90 stars, based on 1 article reviews
tris trizma base - by Bioz Stars, 2026-04
90/100 stars
  Buy from Supplier
concentrated tris base  (Bio-Rad)
98
Bio-Rad concentrated tris base
( a ) The secondary structure of the ladder ribozyme and a schematic showing its function. The ribozyme is shown in red, whilst the substrate strands are shown in black. ( b ) A representative 8% urea PAGE stained with SYBR Gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 19-72 to R3C RNA (total monomer concentration = 1 mM, 10.5 µM substrate, 10.5 µM ribozyme) after a 2 hr reaction at 30°C in 50 <t>mM</t> <t>Tris-HCl</t> pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 0.75:1 (Lys) 19-72 :RNA conditions are shown in blue and red, respectively ( c ) Variation in absorbance at 500 nm as a measure of coacervate formation upon addition of varying ratios of (Lys) 19-72 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( d ) Example fluorescence microscopy image of (Lys) 19-72: RNA condensates at a ratio of 0.75:1 (Lys) 19-72 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( e ) Kinetics of chain elongation in solution (blue, first-order model), and with 0.75:1 (Lys) 19-72 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . A total RNA monomer concentration of 1 mM was achieved by combining 9.5 µM substrate, 1 µM Cy5-tagged substrate and 10.5 µM ribozyme. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( f ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 0.75:1 (Lys) 19-72 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Equivalent data for condensates formed from the shorter (Lys) 5-24 peptide is shown in . Figure 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Figure 1—source data 2. Numerical turbidity data for . Figure 1—source data 3. Unprocessed and uncropped fluorescence microscope image for 0.75:1 (Lys) 19-72 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the accompanying spreadsheet.
Concentrated Tris Base, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/concentrated tris base/product/Bio-Rad
Average 98 stars, based on 1 article reviews
concentrated tris base - by Bioz Stars, 2026-04
98/100 stars
  Buy from Supplier
antigen unmasking solution concentrate  (Vector Laboratories)
96
Vector Laboratories antigen unmasking solution concentrate
( a ) The secondary structure of the ladder ribozyme and a schematic showing its function. The ribozyme is shown in red, whilst the substrate strands are shown in black. ( b ) A representative 8% urea PAGE stained with SYBR Gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 19-72 to R3C RNA (total monomer concentration = 1 mM, 10.5 µM substrate, 10.5 µM ribozyme) after a 2 hr reaction at 30°C in 50 <t>mM</t> <t>Tris-HCl</t> pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 0.75:1 (Lys) 19-72 :RNA conditions are shown in blue and red, respectively ( c ) Variation in absorbance at 500 nm as a measure of coacervate formation upon addition of varying ratios of (Lys) 19-72 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( d ) Example fluorescence microscopy image of (Lys) 19-72: RNA condensates at a ratio of 0.75:1 (Lys) 19-72 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( e ) Kinetics of chain elongation in solution (blue, first-order model), and with 0.75:1 (Lys) 19-72 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . A total RNA monomer concentration of 1 mM was achieved by combining 9.5 µM substrate, 1 µM Cy5-tagged substrate and 10.5 µM ribozyme. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( f ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 0.75:1 (Lys) 19-72 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Equivalent data for condensates formed from the shorter (Lys) 5-24 peptide is shown in . Figure 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Figure 1—source data 2. Numerical turbidity data for . Figure 1—source data 3. Unprocessed and uncropped fluorescence microscope image for 0.75:1 (Lys) 19-72 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the accompanying spreadsheet.
Antigen Unmasking Solution Concentrate, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/antigen unmasking solution concentrate/product/Vector Laboratories
Average 96 stars, based on 1 article reviews
antigen unmasking solution concentrate - by Bioz Stars, 2026-04
96/100 stars
  Buy from Supplier
trizma base  (Millipore)
90
Millipore trizma base
( a ) The secondary structure of the ladder ribozyme and a schematic showing its function. The ribozyme is shown in red, whilst the substrate strands are shown in black. ( b ) A representative 8% urea PAGE stained with SYBR Gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 19-72 to R3C RNA (total monomer concentration = 1 mM, 10.5 µM substrate, 10.5 µM ribozyme) after a 2 hr reaction at 30°C in 50 <t>mM</t> <t>Tris-HCl</t> pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 0.75:1 (Lys) 19-72 :RNA conditions are shown in blue and red, respectively ( c ) Variation in absorbance at 500 nm as a measure of coacervate formation upon addition of varying ratios of (Lys) 19-72 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( d ) Example fluorescence microscopy image of (Lys) 19-72: RNA condensates at a ratio of 0.75:1 (Lys) 19-72 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( e ) Kinetics of chain elongation in solution (blue, first-order model), and with 0.75:1 (Lys) 19-72 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . A total RNA monomer concentration of 1 mM was achieved by combining 9.5 µM substrate, 1 µM Cy5-tagged substrate and 10.5 µM ribozyme. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( f ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 0.75:1 (Lys) 19-72 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Equivalent data for condensates formed from the shorter (Lys) 5-24 peptide is shown in . Figure 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Figure 1—source data 2. Numerical turbidity data for . Figure 1—source data 3. Unprocessed and uncropped fluorescence microscope image for 0.75:1 (Lys) 19-72 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the accompanying spreadsheet.
Trizma Base, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/trizma base/product/Millipore
Average 90 stars, based on 1 article reviews
trizma base - by Bioz Stars, 2026-04
90/100 stars
  Buy from Supplier
tae buffer  (Thermo Fisher)
99
Thermo Fisher tae buffer
( a ) The secondary structure of the ladder ribozyme and a schematic showing its function. The ribozyme is shown in red, whilst the substrate strands are shown in black. ( b ) A representative 8% urea PAGE stained with SYBR Gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 19-72 to R3C RNA (total monomer concentration = 1 mM, 10.5 µM substrate, 10.5 µM ribozyme) after a 2 hr reaction at 30°C in 50 <t>mM</t> <t>Tris-HCl</t> pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 0.75:1 (Lys) 19-72 :RNA conditions are shown in blue and red, respectively ( c ) Variation in absorbance at 500 nm as a measure of coacervate formation upon addition of varying ratios of (Lys) 19-72 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( d ) Example fluorescence microscopy image of (Lys) 19-72: RNA condensates at a ratio of 0.75:1 (Lys) 19-72 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( e ) Kinetics of chain elongation in solution (blue, first-order model), and with 0.75:1 (Lys) 19-72 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . A total RNA monomer concentration of 1 mM was achieved by combining 9.5 µM substrate, 1 µM Cy5-tagged substrate and 10.5 µM ribozyme. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( f ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 0.75:1 (Lys) 19-72 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Equivalent data for condensates formed from the shorter (Lys) 5-24 peptide is shown in . Figure 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Figure 1—source data 2. Numerical turbidity data for . Figure 1—source data 3. Unprocessed and uncropped fluorescence microscope image for 0.75:1 (Lys) 19-72 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the accompanying spreadsheet.
Tae Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/tae buffer/product/Thermo Fisher
Average 99 stars, based on 1 article reviews
tae buffer - by Bioz Stars, 2026-04
99/100 stars
  Buy from Supplier
atp synthesis buffer  (Thermo Fisher)
99
Thermo Fisher atp synthesis buffer
( a ) Representative oxygen flux recording using closed-chamber, high-resolution respirometry. Rat <t>liver</t> <t>mitochondria</t> (0.25 mg protein ml −1 ) were maintained under constant stirring. Oxygen consumption (black trace) and oxygen concentration (grey trace) were measured using the Oxygraph 2 K (Oroboros Instruments, Innsbruck, Austria). Sequential additions of reagents were added to assess respiratory states. ( b ) Mitochondria energised using either glutamate/malate (10 mM/2 mM; black trace) or succinate and rotenone (10 mM/1 μM; grey trace) were treated with oligomycin A (2.5 μg ml −1 ) and FCCP (0.5 μM titration) to assess leak respiration and spare respiratory capacity respectively. ( c ) RCR (respiratory control ratio, calculated as the ratio of State 3/State 4 respiration) for freshly isolated mitochondria and trehalose freeze-thawed mitochondria from the same preparation energised using glutamate/malate (10 mM/2 mM). Statistical significance was calculated using a one-way ANOVA corrected for multiple comparisons using Tukey method (NS P > 0.05; GraphPad Prism). ( d ) RCR for 3 month cryopreserved mitochondria energised using glutamate/malate (10 mM/2 mM). ( e ) Mitochondria (1 mg protein ml −1 ) were incubated in the presence of 1 mM ADP and either glutamate/malate (10 mM/2 mM) or succinate/rotenone (10 mM/1 μM) and total <t>ATP</t> content was measured after 45 minutes. Luminescence was measured in the presence and absence of respiratory inhibitors antimycin A (2.5 μM) and oligomycin A (2.5 μg ml −1 ) to determine background. Data are expressed as means (±s.d.) of at least three independent experiments using mitochondria that had been cryopreserved for between 1 and 3 months. Abbreviations: Mito; mitochondria, Glu; glutamate, Mal; malate, Succ; succinate, FCCP; carbonyl cyanide-p-trifluoromethoxyphenylhydrazone.
Atp Synthesis Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/atp synthesis buffer/product/Thermo Fisher
Average 99 stars, based on 1 article reviews
atp synthesis buffer - by Bioz Stars, 2026-04
99/100 stars
  Buy from Supplier

Image Search Results


( a ) The secondary structure of the ladder ribozyme and a schematic showing its function. The ribozyme is shown in red, whilst the substrate strands are shown in black. ( b ) A representative 8% urea PAGE stained with SYBR Gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 19-72 to R3C RNA (total monomer concentration = 1 mM, 10.5 µM substrate, 10.5 µM ribozyme) after a 2 hr reaction at 30°C in 50 mM Tris-HCl pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 0.75:1 (Lys) 19-72 :RNA conditions are shown in blue and red, respectively ( c ) Variation in absorbance at 500 nm as a measure of coacervate formation upon addition of varying ratios of (Lys) 19-72 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( d ) Example fluorescence microscopy image of (Lys) 19-72: RNA condensates at a ratio of 0.75:1 (Lys) 19-72 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( e ) Kinetics of chain elongation in solution (blue, first-order model), and with 0.75:1 (Lys) 19-72 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . A total RNA monomer concentration of 1 mM was achieved by combining 9.5 µM substrate, 1 µM Cy5-tagged substrate and 10.5 µM ribozyme. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( f ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 0.75:1 (Lys) 19-72 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Equivalent data for condensates formed from the shorter (Lys) 5-24 peptide is shown in . Figure 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Figure 1—source data 2. Numerical turbidity data for . Figure 1—source data 3. Unprocessed and uncropped fluorescence microscope image for 0.75:1 (Lys) 19-72 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the accompanying spreadsheet.

Journal: eLife

Article Title: Ribozyme activity modulates the physical properties of RNA–peptide coacervates

doi: 10.7554/eLife.83543

Figure Lengend Snippet: ( a ) The secondary structure of the ladder ribozyme and a schematic showing its function. The ribozyme is shown in red, whilst the substrate strands are shown in black. ( b ) A representative 8% urea PAGE stained with SYBR Gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 19-72 to R3C RNA (total monomer concentration = 1 mM, 10.5 µM substrate, 10.5 µM ribozyme) after a 2 hr reaction at 30°C in 50 mM Tris-HCl pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 0.75:1 (Lys) 19-72 :RNA conditions are shown in blue and red, respectively ( c ) Variation in absorbance at 500 nm as a measure of coacervate formation upon addition of varying ratios of (Lys) 19-72 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( d ) Example fluorescence microscopy image of (Lys) 19-72: RNA condensates at a ratio of 0.75:1 (Lys) 19-72 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( e ) Kinetics of chain elongation in solution (blue, first-order model), and with 0.75:1 (Lys) 19-72 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . A total RNA monomer concentration of 1 mM was achieved by combining 9.5 µM substrate, 1 µM Cy5-tagged substrate and 10.5 µM ribozyme. Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars are standard deviations. ( f ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 0.75:1 (Lys) 19-72 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Equivalent data for condensates formed from the shorter (Lys) 5-24 peptide is shown in . Figure 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Figure 1—source data 2. Numerical turbidity data for . Figure 1—source data 3. Unprocessed and uncropped fluorescence microscope image for 0.75:1 (Lys) 19-72 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the accompanying spreadsheet.

Article Snippet: Trizma base (Tris; Thermo Fisher Scientific, Waltham, MA), sodium hexametaphosphate ((NaPO 3 ) 6 , 611.77 g/mol; Sigma-Aldrich, St. Louis, MI), formamide (CH 3 NO, 45.04 g/mol; Sigma-Aldrich), ethylenediaminetetracetic acid disodium salt dihydrate (EDTA, C 10 H 14 N 2 Na 2 O 8 ·2H 2 O, 372.24 g/mol; Sigma-Aldrich), magnesium chloride hexahydrate (MgCl 2 ·6H 2 O, 203.30 g/mol; (Merck, Darmstadt, Germany)), sodium hydroxide (NaOH, 39.997 g/mol; VWR, Radnor, PA), sodium chloride (NaCl, 58.44 g/mol; Sigma-Aldrich), urea (CH 4 N 2 O, 60.06 g/mol; Carl Roth, Karlsruhe, Germany), hydrochloric acid (HCl, 37%, 36.46 g/mol) (VWR), boric acid (H 3 BO 3 , 61.83 g/mol; Merck), ammonium persulfate (APS, (NH 4 ) 2 S 2 O 8 , 228.20 g/mol) (VWR), acrylamide (19:1 bisacrylamide; Thermo Fisher Scientific), tetramethylethylendiamine (TEMED, C 6 H 16 N 2 , 116.21 g/mol; Carl Roth), SYBR gold stain (Thermo Fisher Scientific), RNA oligomer length standard (low-range ssRNA ladder) (NEBm Ipswich, MA).

Techniques: Staining, Concentration Assay, Ligation, Fluorescence, Microscopy

The concatenation activity of the ribozyme was determined by reaction at 30, 37, or 45°C and with either equimolar (10.5 µM), twofold (8 µM ribozyme and 16 µM substrate) or fourfold (5 µM ribozyme and 20 µM substrate) substrate concentration relative to the ribozyme. The reaction buffer contained 10 mM MgCl 2 and 50 mM Tris-HCl pH 8.6, and the reaction was stopped after 2 hr. Excess substrate was found to inhibit the formation of long substrate concatenates at lower temperatures. Figure 1—figure supplement 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions.

Journal: eLife

Article Title: Ribozyme activity modulates the physical properties of RNA–peptide coacervates

doi: 10.7554/eLife.83543

Figure Lengend Snippet: The concatenation activity of the ribozyme was determined by reaction at 30, 37, or 45°C and with either equimolar (10.5 µM), twofold (8 µM ribozyme and 16 µM substrate) or fourfold (5 µM ribozyme and 20 µM substrate) substrate concentration relative to the ribozyme. The reaction buffer contained 10 mM MgCl 2 and 50 mM Tris-HCl pH 8.6, and the reaction was stopped after 2 hr. Excess substrate was found to inhibit the formation of long substrate concatenates at lower temperatures. Figure 1—figure supplement 1—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions.

Article Snippet: Trizma base (Tris; Thermo Fisher Scientific, Waltham, MA), sodium hexametaphosphate ((NaPO 3 ) 6 , 611.77 g/mol; Sigma-Aldrich, St. Louis, MI), formamide (CH 3 NO, 45.04 g/mol; Sigma-Aldrich), ethylenediaminetetracetic acid disodium salt dihydrate (EDTA, C 10 H 14 N 2 Na 2 O 8 ·2H 2 O, 372.24 g/mol; Sigma-Aldrich), magnesium chloride hexahydrate (MgCl 2 ·6H 2 O, 203.30 g/mol; (Merck, Darmstadt, Germany)), sodium hydroxide (NaOH, 39.997 g/mol; VWR, Radnor, PA), sodium chloride (NaCl, 58.44 g/mol; Sigma-Aldrich), urea (CH 4 N 2 O, 60.06 g/mol; Carl Roth, Karlsruhe, Germany), hydrochloric acid (HCl, 37%, 36.46 g/mol) (VWR), boric acid (H 3 BO 3 , 61.83 g/mol; Merck), ammonium persulfate (APS, (NH 4 ) 2 S 2 O 8 , 228.20 g/mol) (VWR), acrylamide (19:1 bisacrylamide; Thermo Fisher Scientific), tetramethylethylendiamine (TEMED, C 6 H 16 N 2 , 116.21 g/mol; Carl Roth), SYBR gold stain (Thermo Fisher Scientific), RNA oligomer length standard (low-range ssRNA ladder) (NEBm Ipswich, MA).

Techniques: Activity Assay, Concentration Assay

( a ) A representative 8% urea PAGE gel stained with SYBR gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 5-24 to R3C RNA (total monomer concentration = 1 mM) after a 2 hr reaction at 30°C in 50 mM Tris-HCl pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 1.5:1 (Lys) 5-24 :RNA conditions are shown in blue and red, respectively. ( b ) Variation in absorbance at 500 nm as a proxy for coacervate formation upon addition of varying ratios of (Lys) 5-24 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions with error bars reporting standard deviations. ( c ) Example fluorescence microscopy image of (Lys) 5-24 :RNA condensates at a ratio of 3:1 (Lys) 5-24 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( d ) Kinetics of chain elongation in solution (blue, first-order model), and with 3:1 (Lys) 5-24 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars represent standard deviations ( e ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 3:1 (Lys) 5-24 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Poor sample recovery led to an artificially reduced average substrate length at the t = 30 and t = 60 min time points for the reaction in the presence of (Lys) 5-24 . These points were therefore excluded when fitting data. Figure 1—figure supplement 2—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Content identical to . Figure 1—figure supplement 2—source data 2. Numerical turbidity data. Content identical to . Figure 1—figure supplement 2—source data 3. Unprocessed and uncropped fluorescence microscope image for 3:1 (Lys) 5-24 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—figure supplement 2—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the included spreadsheet. Content identical to .

Journal: eLife

Article Title: Ribozyme activity modulates the physical properties of RNA–peptide coacervates

doi: 10.7554/eLife.83543

Figure Lengend Snippet: ( a ) A representative 8% urea PAGE gel stained with SYBR gold showing the products of the R3C ladder system in solution and varying ratios of (Lys) 5-24 to R3C RNA (total monomer concentration = 1 mM) after a 2 hr reaction at 30°C in 50 mM Tris-HCl pH 8.6 and 10 mM MgCl 2 . The integrated lane profiles of the solution and 1.5:1 (Lys) 5-24 :RNA conditions are shown in blue and red, respectively. ( b ) Variation in absorbance at 500 nm as a proxy for coacervate formation upon addition of varying ratios of (Lys) 5-24 to the E L RNA after ligation for 3 hr at 30°C. Data points are an average of n = 3 independent replicates assembled from the same stock solutions with error bars reporting standard deviations. ( c ) Example fluorescence microscopy image of (Lys) 5-24 :RNA condensates at a ratio of 3:1 (Lys) 5-24 :RNA, imaged using 10% Cy5-tagged substrate strand. Scale bar = 20 μm. ( d ) Kinetics of chain elongation in solution (blue, first-order model), and with 3:1 (Lys) 5-24 :RNA (red, second-order model) at 30°C, pH 8.6, and 10 mM MgCl 2 . Data points are an average of n = 3 independent replicates assembled from the same stock solutions. Error bars represent standard deviations ( e ) Chain extension rate constants for the R3C ladder ribozyme in solution (blue, first-order model) and in the presence of 3:1 (Lys) 5-24 :RNA (red, second-order model). Error bars are the standard errors for each parameter computed during non-linear regression. Poor sample recovery led to an artificially reduced average substrate length at the t = 30 and t = 60 min time points for the reaction in the presence of (Lys) 5-24 . These points were therefore excluded when fitting data. Figure 1—figure supplement 2—source data 1. Unedited and uncropped gel image for , and labelled image showing key bands and conditions. Content identical to . Figure 1—figure supplement 2—source data 2. Numerical turbidity data. Content identical to . Figure 1—figure supplement 2—source data 3. Unprocessed and uncropped fluorescence microscope image for 3:1 (Lys) 5-24 :RNA condensates imaged using 10% Cy5-tagged substrate strand. Figure 1—figure supplement 2—source data 4. Unedited and uncropped gel images for ribozyme kinetics . Lane identities are listed in the included spreadsheet. Content identical to .

Article Snippet: Trizma base (Tris; Thermo Fisher Scientific, Waltham, MA), sodium hexametaphosphate ((NaPO 3 ) 6 , 611.77 g/mol; Sigma-Aldrich, St. Louis, MI), formamide (CH 3 NO, 45.04 g/mol; Sigma-Aldrich), ethylenediaminetetracetic acid disodium salt dihydrate (EDTA, C 10 H 14 N 2 Na 2 O 8 ·2H 2 O, 372.24 g/mol; Sigma-Aldrich), magnesium chloride hexahydrate (MgCl 2 ·6H 2 O, 203.30 g/mol; (Merck, Darmstadt, Germany)), sodium hydroxide (NaOH, 39.997 g/mol; VWR, Radnor, PA), sodium chloride (NaCl, 58.44 g/mol; Sigma-Aldrich), urea (CH 4 N 2 O, 60.06 g/mol; Carl Roth, Karlsruhe, Germany), hydrochloric acid (HCl, 37%, 36.46 g/mol) (VWR), boric acid (H 3 BO 3 , 61.83 g/mol; Merck), ammonium persulfate (APS, (NH 4 ) 2 S 2 O 8 , 228.20 g/mol) (VWR), acrylamide (19:1 bisacrylamide; Thermo Fisher Scientific), tetramethylethylendiamine (TEMED, C 6 H 16 N 2 , 116.21 g/mol; Carl Roth), SYBR gold stain (Thermo Fisher Scientific), RNA oligomer length standard (low-range ssRNA ladder) (NEBm Ipswich, MA).

Techniques: Staining, Concentration Assay, Ligation, Fluorescence, Microscopy

Experiments were performed in solution at 30°C in 50 mM Tris-HCl pH 8.6, 10 mM MgCl 2 , and with the addition of either 1.5:1 (Lys) 5-24 :RNA or 0.75:1 (Lys) 19-72 :RNA. The reaction run using 10% Cy5-tagged substrate strand, and the reaction product were visualised on an 8% urea PAGE. The displayed gels correspond to the kinetic plots shown in and . Figure 1—figure supplement 3—source data 1. Source data contains unedited and uncropped gel images used to estimate yields, as well as labelled images showing key bands and conditions and is identical to .

Journal: eLife

Article Title: Ribozyme activity modulates the physical properties of RNA–peptide coacervates

doi: 10.7554/eLife.83543

Figure Lengend Snippet: Experiments were performed in solution at 30°C in 50 mM Tris-HCl pH 8.6, 10 mM MgCl 2 , and with the addition of either 1.5:1 (Lys) 5-24 :RNA or 0.75:1 (Lys) 19-72 :RNA. The reaction run using 10% Cy5-tagged substrate strand, and the reaction product were visualised on an 8% urea PAGE. The displayed gels correspond to the kinetic plots shown in and . Figure 1—figure supplement 3—source data 1. Source data contains unedited and uncropped gel images used to estimate yields, as well as labelled images showing key bands and conditions and is identical to .

Article Snippet: Trizma base (Tris; Thermo Fisher Scientific, Waltham, MA), sodium hexametaphosphate ((NaPO 3 ) 6 , 611.77 g/mol; Sigma-Aldrich, St. Louis, MI), formamide (CH 3 NO, 45.04 g/mol; Sigma-Aldrich), ethylenediaminetetracetic acid disodium salt dihydrate (EDTA, C 10 H 14 N 2 Na 2 O 8 ·2H 2 O, 372.24 g/mol; Sigma-Aldrich), magnesium chloride hexahydrate (MgCl 2 ·6H 2 O, 203.30 g/mol; (Merck, Darmstadt, Germany)), sodium hydroxide (NaOH, 39.997 g/mol; VWR, Radnor, PA), sodium chloride (NaCl, 58.44 g/mol; Sigma-Aldrich), urea (CH 4 N 2 O, 60.06 g/mol; Carl Roth, Karlsruhe, Germany), hydrochloric acid (HCl, 37%, 36.46 g/mol) (VWR), boric acid (H 3 BO 3 , 61.83 g/mol; Merck), ammonium persulfate (APS, (NH 4 ) 2 S 2 O 8 , 228.20 g/mol) (VWR), acrylamide (19:1 bisacrylamide; Thermo Fisher Scientific), tetramethylethylendiamine (TEMED, C 6 H 16 N 2 , 116.21 g/mol; Carl Roth), SYBR gold stain (Thermo Fisher Scientific), RNA oligomer length standard (low-range ssRNA ladder) (NEBm Ipswich, MA).

Techniques:

Ribozyme assays were performed at 45 °C in solution and in the presence of poly(L-lysine) (0.75:1 Lys 19-72 :RNA or 3:1 Lys 5-24 :RNA). The ribozyme reaction buffer contained 10 mM MgCl 2 and 50 mM Tris-HCl pH 8.6. The reaction was stopped after 3 hr and the extracted RNA was digested with RNase R, leaving only circular products. Circular bands are marked with an asterisk. The formation of circular products is observed in solution but is suppressed in the presence of poly(L-lysine). Figure 1—figure supplement 5—source data 1. Unedited and uncropped gel image, and labelled image showing key bands and conditions.

Journal: eLife

Article Title: Ribozyme activity modulates the physical properties of RNA–peptide coacervates

doi: 10.7554/eLife.83543

Figure Lengend Snippet: Ribozyme assays were performed at 45 °C in solution and in the presence of poly(L-lysine) (0.75:1 Lys 19-72 :RNA or 3:1 Lys 5-24 :RNA). The ribozyme reaction buffer contained 10 mM MgCl 2 and 50 mM Tris-HCl pH 8.6. The reaction was stopped after 3 hr and the extracted RNA was digested with RNase R, leaving only circular products. Circular bands are marked with an asterisk. The formation of circular products is observed in solution but is suppressed in the presence of poly(L-lysine). Figure 1—figure supplement 5—source data 1. Unedited and uncropped gel image, and labelled image showing key bands and conditions.

Article Snippet: Trizma base (Tris; Thermo Fisher Scientific, Waltham, MA), sodium hexametaphosphate ((NaPO 3 ) 6 , 611.77 g/mol; Sigma-Aldrich, St. Louis, MI), formamide (CH 3 NO, 45.04 g/mol; Sigma-Aldrich), ethylenediaminetetracetic acid disodium salt dihydrate (EDTA, C 10 H 14 N 2 Na 2 O 8 ·2H 2 O, 372.24 g/mol; Sigma-Aldrich), magnesium chloride hexahydrate (MgCl 2 ·6H 2 O, 203.30 g/mol; (Merck, Darmstadt, Germany)), sodium hydroxide (NaOH, 39.997 g/mol; VWR, Radnor, PA), sodium chloride (NaCl, 58.44 g/mol; Sigma-Aldrich), urea (CH 4 N 2 O, 60.06 g/mol; Carl Roth, Karlsruhe, Germany), hydrochloric acid (HCl, 37%, 36.46 g/mol) (VWR), boric acid (H 3 BO 3 , 61.83 g/mol; Merck), ammonium persulfate (APS, (NH 4 ) 2 S 2 O 8 , 228.20 g/mol) (VWR), acrylamide (19:1 bisacrylamide; Thermo Fisher Scientific), tetramethylethylendiamine (TEMED, C 6 H 16 N 2 , 116.21 g/mol; Carl Roth), SYBR gold stain (Thermo Fisher Scientific), RNA oligomer length standard (low-range ssRNA ladder) (NEBm Ipswich, MA).

Techniques:

The activity of the active and inactive ligase ribozyme variants was tested in solution and in the presence of poly(L-lysine) (0.75:1 Lys 19-72 :RNA or 3:1 Lys 5-24 :RNA). The ribozyme reaction buffer contained 10 mM MgCl 2 and 50 mM Tris-HCl pH 8.6. The reaction was stopped after 2 hr. No ligation activity was detected in the presence of the inactive ribozyme in all conditions tested. Figure 2—figure supplement 1—source data 1. Unedited and uncropped gel image, and labelled image showing key bands and conditions.

Journal: eLife

Article Title: Ribozyme activity modulates the physical properties of RNA–peptide coacervates

doi: 10.7554/eLife.83543

Figure Lengend Snippet: The activity of the active and inactive ligase ribozyme variants was tested in solution and in the presence of poly(L-lysine) (0.75:1 Lys 19-72 :RNA or 3:1 Lys 5-24 :RNA). The ribozyme reaction buffer contained 10 mM MgCl 2 and 50 mM Tris-HCl pH 8.6. The reaction was stopped after 2 hr. No ligation activity was detected in the presence of the inactive ribozyme in all conditions tested. Figure 2—figure supplement 1—source data 1. Unedited and uncropped gel image, and labelled image showing key bands and conditions.

Article Snippet: Trizma base (Tris; Thermo Fisher Scientific, Waltham, MA), sodium hexametaphosphate ((NaPO 3 ) 6 , 611.77 g/mol; Sigma-Aldrich, St. Louis, MI), formamide (CH 3 NO, 45.04 g/mol; Sigma-Aldrich), ethylenediaminetetracetic acid disodium salt dihydrate (EDTA, C 10 H 14 N 2 Na 2 O 8 ·2H 2 O, 372.24 g/mol; Sigma-Aldrich), magnesium chloride hexahydrate (MgCl 2 ·6H 2 O, 203.30 g/mol; (Merck, Darmstadt, Germany)), sodium hydroxide (NaOH, 39.997 g/mol; VWR, Radnor, PA), sodium chloride (NaCl, 58.44 g/mol; Sigma-Aldrich), urea (CH 4 N 2 O, 60.06 g/mol; Carl Roth, Karlsruhe, Germany), hydrochloric acid (HCl, 37%, 36.46 g/mol) (VWR), boric acid (H 3 BO 3 , 61.83 g/mol; Merck), ammonium persulfate (APS, (NH 4 ) 2 S 2 O 8 , 228.20 g/mol) (VWR), acrylamide (19:1 bisacrylamide; Thermo Fisher Scientific), tetramethylethylendiamine (TEMED, C 6 H 16 N 2 , 116.21 g/mol; Carl Roth), SYBR gold stain (Thermo Fisher Scientific), RNA oligomer length standard (low-range ssRNA ladder) (NEBm Ipswich, MA).

Techniques: Activity Assay, Ligation

( a ) Representative oxygen flux recording using closed-chamber, high-resolution respirometry. Rat liver mitochondria (0.25 mg protein ml −1 ) were maintained under constant stirring. Oxygen consumption (black trace) and oxygen concentration (grey trace) were measured using the Oxygraph 2 K (Oroboros Instruments, Innsbruck, Austria). Sequential additions of reagents were added to assess respiratory states. ( b ) Mitochondria energised using either glutamate/malate (10 mM/2 mM; black trace) or succinate and rotenone (10 mM/1 μM; grey trace) were treated with oligomycin A (2.5 μg ml −1 ) and FCCP (0.5 μM titration) to assess leak respiration and spare respiratory capacity respectively. ( c ) RCR (respiratory control ratio, calculated as the ratio of State 3/State 4 respiration) for freshly isolated mitochondria and trehalose freeze-thawed mitochondria from the same preparation energised using glutamate/malate (10 mM/2 mM). Statistical significance was calculated using a one-way ANOVA corrected for multiple comparisons using Tukey method (NS P > 0.05; GraphPad Prism). ( d ) RCR for 3 month cryopreserved mitochondria energised using glutamate/malate (10 mM/2 mM). ( e ) Mitochondria (1 mg protein ml −1 ) were incubated in the presence of 1 mM ADP and either glutamate/malate (10 mM/2 mM) or succinate/rotenone (10 mM/1 μM) and total ATP content was measured after 45 minutes. Luminescence was measured in the presence and absence of respiratory inhibitors antimycin A (2.5 μM) and oligomycin A (2.5 μg ml −1 ) to determine background. Data are expressed as means (±s.d.) of at least three independent experiments using mitochondria that had been cryopreserved for between 1 and 3 months. Abbreviations: Mito; mitochondria, Glu; glutamate, Mal; malate, Succ; succinate, FCCP; carbonyl cyanide-p-trifluoromethoxyphenylhydrazone.

Journal: Scientific Reports

Article Title: Identification of ER-000444793, a Cyclophilin D-independent inhibitor of mitochondrial permeability transition, using a high-throughput screen in cryopreserved mitochondria

doi: 10.1038/srep37798

Figure Lengend Snippet: ( a ) Representative oxygen flux recording using closed-chamber, high-resolution respirometry. Rat liver mitochondria (0.25 mg protein ml −1 ) were maintained under constant stirring. Oxygen consumption (black trace) and oxygen concentration (grey trace) were measured using the Oxygraph 2 K (Oroboros Instruments, Innsbruck, Austria). Sequential additions of reagents were added to assess respiratory states. ( b ) Mitochondria energised using either glutamate/malate (10 mM/2 mM; black trace) or succinate and rotenone (10 mM/1 μM; grey trace) were treated with oligomycin A (2.5 μg ml −1 ) and FCCP (0.5 μM titration) to assess leak respiration and spare respiratory capacity respectively. ( c ) RCR (respiratory control ratio, calculated as the ratio of State 3/State 4 respiration) for freshly isolated mitochondria and trehalose freeze-thawed mitochondria from the same preparation energised using glutamate/malate (10 mM/2 mM). Statistical significance was calculated using a one-way ANOVA corrected for multiple comparisons using Tukey method (NS P > 0.05; GraphPad Prism). ( d ) RCR for 3 month cryopreserved mitochondria energised using glutamate/malate (10 mM/2 mM). ( e ) Mitochondria (1 mg protein ml −1 ) were incubated in the presence of 1 mM ADP and either glutamate/malate (10 mM/2 mM) or succinate/rotenone (10 mM/1 μM) and total ATP content was measured after 45 minutes. Luminescence was measured in the presence and absence of respiratory inhibitors antimycin A (2.5 μM) and oligomycin A (2.5 μg ml −1 ) to determine background. Data are expressed as means (±s.d.) of at least three independent experiments using mitochondria that had been cryopreserved for between 1 and 3 months. Abbreviations: Mito; mitochondria, Glu; glutamate, Mal; malate, Succ; succinate, FCCP; carbonyl cyanide-p-trifluoromethoxyphenylhydrazone.

Article Snippet: Mitochondria (1 mg protein ml −1 final concentration) were re-suspended in ATP synthesis buffer (20 mM Tris base 0.6 M sorbitol, 15 mM potassium phosphate monobasic, 10 mM magnesium sulphate, 2.5 mg ml −1 essentially fatty acid free BSA, final pH 7.4) containing either 10 mM L-glutamic acid, monosodium salt; 2 mM L-malic acid sodium salt or 10 mM succinate disodium salt; 1 μM rotenone and mitochondrial suspension (20 μl) was dispensed into a clear bottom black-walled 384 well plate using a Multidrop Combi Reagent Dispenser (Thermo Scientific, Rockford, IL).

Techniques: Concentration Assay, Titration, Control, Isolation, Incubation

( a,b ) Rat liver mitochondria (1 mg protein ml −1 ) were incubated with compound for 10 minutes in the presence of either glutamate (10 mM) and malate (2 mM) or succinate (10 mM) and rotenone (1 μM). ADP (5 mM) was added and incubated for 45 minutes. Mitochondria were lysed and total ATP quantified by luminescence using CellTiter Glo reagent (Promega, Madison, WI). No effect of either ER-000444793, CsA or SfA was observed under either respiratory substrate condition. Results are expressed as % inhibition of ATP, synthesis normalised to DMSO (0% inhibition) and antimycin A (2.5 μM) plus oligomycin A (2.5 μg ml −1 ; 100% inhibition). Data are expressed as means (± s.d.) of at least three independent experiments. ( c ) Assessment of immunosuppression and cellular toxicity in Jurkat NFAT-reporter cells. Jurkat cells were incubated with compound for 45 minutes prior to the addition of ionomycin (0.5 μM) and PMA (50 nM). NFAT reporter activity was assessed after 20 hours treatment with either ER-000444793, CsA or SfA and quantified by luminescence using Bright Glo reagent (Promega, Madison, WI). Data are expressed as % inhibition, normalised to DMSO (0% inhibition) and CsA (5 μM; 100% inhibition). ( d ) Toxicity of ionomycin/PMA treatment was assessed in Jurkat cells using Alamar blue after 20 hours incubation. No significant effect was observed with treatment. Data are expressed as fluorescence intensity and statistical significance calculated using paired two-tailed student’s t -test using GraphPad Prism (NS P > 0.05). ( e ) Jurkat cells were incubated with compound for 20 hours and toxicity assessed using Alamar Blue. Data are expressed as fluorescence intensity and compared using multiple t-tests comparing treatments to DMSO, corrected for multiple comparisons using Holm-Sidak method , P < 0.05 defined as being statistically significant. All data are expressed as means (± s.d.) of three independent experiments. Abbreviations: CsA; cyclosporin A, SfA; sanglifehrin A, Cmp; compound, O.D; optical density, NS; not significant, PMA; phorbol 12-myristate.

Journal: Scientific Reports

Article Title: Identification of ER-000444793, a Cyclophilin D-independent inhibitor of mitochondrial permeability transition, using a high-throughput screen in cryopreserved mitochondria

doi: 10.1038/srep37798

Figure Lengend Snippet: ( a,b ) Rat liver mitochondria (1 mg protein ml −1 ) were incubated with compound for 10 minutes in the presence of either glutamate (10 mM) and malate (2 mM) or succinate (10 mM) and rotenone (1 μM). ADP (5 mM) was added and incubated for 45 minutes. Mitochondria were lysed and total ATP quantified by luminescence using CellTiter Glo reagent (Promega, Madison, WI). No effect of either ER-000444793, CsA or SfA was observed under either respiratory substrate condition. Results are expressed as % inhibition of ATP, synthesis normalised to DMSO (0% inhibition) and antimycin A (2.5 μM) plus oligomycin A (2.5 μg ml −1 ; 100% inhibition). Data are expressed as means (± s.d.) of at least three independent experiments. ( c ) Assessment of immunosuppression and cellular toxicity in Jurkat NFAT-reporter cells. Jurkat cells were incubated with compound for 45 minutes prior to the addition of ionomycin (0.5 μM) and PMA (50 nM). NFAT reporter activity was assessed after 20 hours treatment with either ER-000444793, CsA or SfA and quantified by luminescence using Bright Glo reagent (Promega, Madison, WI). Data are expressed as % inhibition, normalised to DMSO (0% inhibition) and CsA (5 μM; 100% inhibition). ( d ) Toxicity of ionomycin/PMA treatment was assessed in Jurkat cells using Alamar blue after 20 hours incubation. No significant effect was observed with treatment. Data are expressed as fluorescence intensity and statistical significance calculated using paired two-tailed student’s t -test using GraphPad Prism (NS P > 0.05). ( e ) Jurkat cells were incubated with compound for 20 hours and toxicity assessed using Alamar Blue. Data are expressed as fluorescence intensity and compared using multiple t-tests comparing treatments to DMSO, corrected for multiple comparisons using Holm-Sidak method , P < 0.05 defined as being statistically significant. All data are expressed as means (± s.d.) of three independent experiments. Abbreviations: CsA; cyclosporin A, SfA; sanglifehrin A, Cmp; compound, O.D; optical density, NS; not significant, PMA; phorbol 12-myristate.

Article Snippet: Mitochondria (1 mg protein ml −1 final concentration) were re-suspended in ATP synthesis buffer (20 mM Tris base 0.6 M sorbitol, 15 mM potassium phosphate monobasic, 10 mM magnesium sulphate, 2.5 mg ml −1 essentially fatty acid free BSA, final pH 7.4) containing either 10 mM L-glutamic acid, monosodium salt; 2 mM L-malic acid sodium salt or 10 mM succinate disodium salt; 1 μM rotenone and mitochondrial suspension (20 μl) was dispensed into a clear bottom black-walled 384 well plate using a Multidrop Combi Reagent Dispenser (Thermo Scientific, Rockford, IL).

Techniques: Incubation, Inhibition, Activity Assay, Fluorescence, Two Tailed Test