20 mm tris Search Results


tris-buffered saline (tbs: 20 mm tris, 150 mm nacl)  (Matreya LLC)
90
Matreya LLC tris-buffered saline (tbs: 20 mm tris, 150 mm nacl)
Tris Buffered Saline (Tbs: 20 Mm Tris, 150 Mm Nacl), supplied by Matreya LLC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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buffer tr.a (50 mm tris-hcl, 30 mm nacl, ph 8.0)  (Constant Systems Ltd)
90
Constant Systems Ltd buffer tr.a (50 mm tris-hcl, 30 mm nacl, ph 8.0)
Buffer Tr.A (50 Mm Tris Hcl, 30 Mm Nacl, Ph 8.0), supplied by Constant Systems Ltd, 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/buffer tr.a (50 mm tris-hcl, 30 mm nacl, ph 8.0)/product/Constant Systems Ltd
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buffer tr.a (50 mm tris-hcl, 30 mm nacl, ph 8.0) - by Bioz Stars, 2026-02
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tris-hcl buffer (ph 8.0) containing nacl  (Spectrum Labs)
90
Spectrum Labs tris-hcl buffer (ph 8.0) containing nacl
Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
Tris Hcl Buffer (Ph 8.0) Containing Nacl, supplied by Spectrum Labs, 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-hcl buffer (ph 8.0) containing nacl/product/Spectrum Labs
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20 mm tris-cl (ph=8.0) in 15% acetonitrile  (Avantor)
90
Avantor 20 mm tris-cl (ph=8.0) in 15% acetonitrile
Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
20 Mm Tris Cl (Ph=8.0) In 15% Acetonitrile, supplied by Avantor, 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/20 mm tris-cl (ph=8.0) in 15% acetonitrile/product/Avantor
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20 mm tris-cl (ph=8.0) in 15% acetonitrile - by Bioz Stars, 2026-02
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blocking reagent 2% block and 0.2% sodium dodecylsulphate in maleic acid, ph 6.0  (Boehringer Mannheim)
90
Boehringer Mannheim blocking reagent 2% block and 0.2% sodium dodecylsulphate in maleic acid, ph 6.0
Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
Blocking Reagent 2% Block And 0.2% Sodium Dodecylsulphate In Maleic Acid, Ph 6.0, supplied by Boehringer Mannheim, 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/blocking reagent 2% block and 0.2% sodium dodecylsulphate in maleic acid, ph 6.0/product/Boehringer Mannheim
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blocking reagent 2% block and 0.2% sodium dodecylsulphate in maleic acid, ph 6.0 - by Bioz Stars, 2026-02
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transferbuffer [50 mm tris, 40 mm glycine, 20% (v/v) methanol, ph 8.6]  (Schuell GmbH)
90
Schuell GmbH transferbuffer [50 mm tris, 40 mm glycine, 20% (v/v) methanol, ph 8.6]
Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
Transferbuffer [50 Mm Tris, 40 Mm Glycine, 20% (V/V) Methanol, Ph 8.6], supplied by Schuell 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/transferbuffer [50 mm tris, 40 mm glycine, 20% (v/v) methanol, ph 8.6]/product/Schuell GmbH
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transferbuffer [50 mm tris, 40 mm glycine, 20% (v/v) methanol, ph 8.6] - by Bioz Stars, 2026-02
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20 mm tris buffer  (Bruker Corporation)
90
Bruker Corporation 20 mm tris buffer
Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
20 Mm Tris Buffer, supplied by Bruker Corporation, 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/20 mm tris buffer/product/Bruker Corporation
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20 mm tris buffer - by Bioz Stars, 2026-02
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20 mg/ml dermatan sulfate solution in 50 mm tris-hcl/60 mm sodium acetate buffer, ph 8  (Seikagaku corporation)
90
Seikagaku corporation 20 mg/ml dermatan sulfate solution in 50 mm tris-hcl/60 mm sodium acetate buffer, ph 8
Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
20 Mg/Ml Dermatan Sulfate Solution In 50 Mm Tris Hcl/60 Mm Sodium Acetate Buffer, Ph 8, supplied by Seikagaku corporation, 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/20 mg/ml dermatan sulfate solution in 50 mm tris-hcl/60 mm sodium acetate buffer, ph 8/product/Seikagaku corporation
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20 mg/ml dermatan sulfate solution in 50 mm tris-hcl/60 mm sodium acetate buffer, ph 8 - by Bioz Stars, 2026-02
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hbsg buffer (20 mm hepes, 150 mm nacl, 20 % glycerol, ph 7.8)  (Nacalai)
90
Nacalai hbsg buffer (20 mm hepes, 150 mm nacl, 20 % glycerol, ph 7.8)
Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
Hbsg Buffer (20 Mm Hepes, 150 Mm Nacl, 20 % Glycerol, Ph 7.8), supplied by Nacalai, 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/hbsg buffer (20 mm hepes, 150 mm nacl, 20 % glycerol, ph 7.8)/product/Nacalai
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hbsg buffer (20 mm hepes, 150 mm nacl, 20 % glycerol, ph 7.8) - by Bioz Stars, 2026-02
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lysis buffer 20-mm tris  (Merck KGaA)
90
Merck KGaA lysis buffer 20-mm tris
Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
Lysis Buffer 20 Mm Tris, supplied by Merck KGaA, 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/lysis buffer 20-mm tris/product/Merck KGaA
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lysis buffer 20-mm tris - by Bioz Stars, 2026-02
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tbs buffer (50 mm tris, 150 mm nacl, ph 7.4) + 0.2% tween-20  (Applichem inc)
90
Applichem inc tbs buffer (50 mm tris, 150 mm nacl, ph 7.4) + 0.2% tween-20
Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
Tbs Buffer (50 Mm Tris, 150 Mm Nacl, Ph 7.4) + 0.2% Tween 20, supplied by Applichem inc, 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/tbs buffer (50 mm tris, 150 mm nacl, ph 7.4) + 0.2% tween-20/product/Applichem inc
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tbs buffer (50 mm tris, 150 mm nacl, ph 7.4) + 0.2% tween-20 - by Bioz Stars, 2026-02
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recombinant his 6 -tagged smeldhp  (Hampton Research Corp)
90
Hampton Research Corp recombinant his 6 -tagged smeldhp
(a) SDS–PAGE analysis of each purification step of the dihydropyrimidinase from S. meliloti CECT4114. Lanes 1 and 2, supernatant and pellet of the resuspended crude extract after cell sonication; lane 3, eluate after adding the sonicated supernatant to the metal-affinity column; lane 4, flowthrough after washing the metal-affinity column with buffer; lane 5, purified <t>SmelDhp</t> (4.25 µg); lane 6, low-molecular-weight markers (kDa). (b) Size-exclusion chromatography of the purified SmelDhp. The enzyme showed a homotetrameric native structure.
Recombinant His 6 Tagged Smeldhp, supplied by Hampton Research Corp, 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/recombinant his 6 -tagged smeldhp/product/Hampton Research Corp
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recombinant his 6 -tagged smeldhp - by Bioz Stars, 2026-02
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Image Search Results


Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.

Journal: The Journal of Biological Chemistry

Article Title: Modeling protease-sensitive human pancreatic lipase mutations in the mouse ortholog

doi: 10.1016/j.jbc.2024.107763

Figure Lengend Snippet: Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.

Article Snippet: Fractions containing pure PNLIP protein were pooled and dialyzed against 3 × 1000 ml of 50 mM Tris-HCl buffer (pH 8.0) containing 100 mM NaCl using Spectra/Por Float-A-Lyzer Dialysis devices (Spectrum Labs) with molecular weight cut-off of 10 kDa.

Techniques: Purification, Mutagenesis, Concentration Assay, Incubation, SDS Page, Staining

Effect of mouse chymotrypsin C (CTRC) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRC in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

Journal: The Journal of Biological Chemistry

Article Title: Modeling protease-sensitive human pancreatic lipase mutations in the mouse ortholog

doi: 10.1016/j.jbc.2024.107763

Figure Lengend Snippet: Effect of mouse chymotrypsin C (CTRC) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRC in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

Article Snippet: Fractions containing pure PNLIP protein were pooled and dialyzed against 3 × 1000 ml of 50 mM Tris-HCl buffer (pH 8.0) containing 100 mM NaCl using Spectra/Por Float-A-Lyzer Dialysis devices (Spectrum Labs) with molecular weight cut-off of 10 kDa.

Techniques: Purification, Mutagenesis, Concentration Assay, Incubation, SDS Page, Staining

Effect of mouse chymotrypsin B1 (CTRB1) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRB1 in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

Journal: The Journal of Biological Chemistry

Article Title: Modeling protease-sensitive human pancreatic lipase mutations in the mouse ortholog

doi: 10.1016/j.jbc.2024.107763

Figure Lengend Snippet: Effect of mouse chymotrypsin B1 (CTRB1) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRB1 in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

Article Snippet: Fractions containing pure PNLIP protein were pooled and dialyzed against 3 × 1000 ml of 50 mM Tris-HCl buffer (pH 8.0) containing 100 mM NaCl using Spectra/Por Float-A-Lyzer Dialysis devices (Spectrum Labs) with molecular weight cut-off of 10 kDa.

Techniques: Purification, Mutagenesis, Concentration Assay, Incubation, SDS Page, Staining

Effect of the T341K mutation on the degradation of mouse PNLIP variants by mouse T7 trypsin. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.

Journal: The Journal of Biological Chemistry

Article Title: Modeling protease-sensitive human pancreatic lipase mutations in the mouse ortholog

doi: 10.1016/j.jbc.2024.107763

Figure Lengend Snippet: Effect of the T341K mutation on the degradation of mouse PNLIP variants by mouse T7 trypsin. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.

Article Snippet: Fractions containing pure PNLIP protein were pooled and dialyzed against 3 × 1000 ml of 50 mM Tris-HCl buffer (pH 8.0) containing 100 mM NaCl using Spectra/Por Float-A-Lyzer Dialysis devices (Spectrum Labs) with molecular weight cut-off of 10 kDa.

Techniques: Mutagenesis, Purification, Concentration Assay, Incubation, SDS Page, Staining

(a) SDS–PAGE analysis of each purification step of the dihydropyrimidinase from S. meliloti CECT4114. Lanes 1 and 2, supernatant and pellet of the resuspended crude extract after cell sonication; lane 3, eluate after adding the sonicated supernatant to the metal-affinity column; lane 4, flowthrough after washing the metal-affinity column with buffer; lane 5, purified SmelDhp (4.25 µg); lane 6, low-molecular-weight markers (kDa). (b) Size-exclusion chromatography of the purified SmelDhp. The enzyme showed a homotetrameric native structure.

Journal:

Article Title: Crystallization and preliminary crystallographic studies of the recombinant dihydropyrimidinase from Sinorhizobium meliloti CECT4114

doi: 10.1107/S1744309106045362

Figure Lengend Snippet: (a) SDS–PAGE analysis of each purification step of the dihydropyrimidinase from S. meliloti CECT4114. Lanes 1 and 2, supernatant and pellet of the resuspended crude extract after cell sonication; lane 3, eluate after adding the sonicated supernatant to the metal-affinity column; lane 4, flowthrough after washing the metal-affinity column with buffer; lane 5, purified SmelDhp (4.25 µg); lane 6, low-molecular-weight markers (kDa). (b) Size-exclusion chromatography of the purified SmelDhp. The enzyme showed a homotetrameric native structure.

Article Snippet: Recombinant His 6 -tagged SmelDhp at a concentration of 18 mg ml −1 in 20 m M Tris–HCl pH 8.0 and 0.5 m M ZnCl 2 was used to perform initial crystallization screening with Hampton Research Crystal Screen I (based on the sparse-matrix method; Jancarik & Kim, 1991 ▶ ) at 293 K. The hanging-drop vapour-diffusion method was used, with drops made by mixing equal volumes (2 μl) of 18 mg ml −1 enzyme solution and reservoir solution, which were suspended over 1.0 ml reservoir.

Techniques: SDS Page, Purification, Sonication, Affinity Column, Molecular Weight, Size-exclusion Chromatography

(a) Crystals of recombinant SmelDhp grown by the vapour-diffusion method. (b) and (c) Crystals of recombinant SmelDhp grown in 0.3 mm inner diameter capillaries in the presence of 0.1%(w/v) agarose by the counter-diffusion technique.

Journal:

Article Title: Crystallization and preliminary crystallographic studies of the recombinant dihydropyrimidinase from Sinorhizobium meliloti CECT4114

doi: 10.1107/S1744309106045362

Figure Lengend Snippet: (a) Crystals of recombinant SmelDhp grown by the vapour-diffusion method. (b) and (c) Crystals of recombinant SmelDhp grown in 0.3 mm inner diameter capillaries in the presence of 0.1%(w/v) agarose by the counter-diffusion technique.

Article Snippet: Recombinant His 6 -tagged SmelDhp at a concentration of 18 mg ml −1 in 20 m M Tris–HCl pH 8.0 and 0.5 m M ZnCl 2 was used to perform initial crystallization screening with Hampton Research Crystal Screen I (based on the sparse-matrix method; Jancarik & Kim, 1991 ▶ ) at 293 K. The hanging-drop vapour-diffusion method was used, with drops made by mixing equal volumes (2 μl) of 18 mg ml −1 enzyme solution and reservoir solution, which were suspended over 1.0 ml reservoir.

Techniques: Recombinant, Diffusion-based Assay