Review



buffer 2 mm tcep  (Selleck Chemicals)


Bioz Verified Symbol Selleck Chemicals is a verified supplier
Bioz Manufacturer Symbol Selleck Chemicals manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93

    Structured Review

    Selleck Chemicals buffer 2 mm tcep
    Buffer 2 Mm Tcep, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 93/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/buffer+2+mm+tcep/Glycerol/pm41931608-290-36-41
    Average 93 stars, based on 53 article reviews
    buffer 2 mm tcep - by Bioz Stars, 2026-09
    93/100 stars

    Images

    Related Articles

    Protein Purification:

    Article Title: Cryo-EM structures of human P2X2/3 heteromer channel reveal the structural basis of ligand selectivity
    Article Snippet: .. For P2X2 or P2X3 protein purification, 500-ml cell pellets were resuspended in 25 ml of hypotonic buffer (25 mM Hepes, pH 7.5, 150 mM NaCl, 5 mM CaCl 2 , 2 mM TCEP, and 10% glycerol) supplemented with a protease inhibitor cocktail (Selleck) for 30 min with gentle agitation. ..

    Article Title: Cryo-EM structures of human P2X2/3 heteromer channel reveal the structural basis of ligand selectivity.
    Article Snippet: .. For P2X2 or P2X3 protein purification, 500- ml cell pellets were resuspended in 25 ml of hypotonic buffer (25 mM Hepes, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 2 mM TCEP, and 10% glycerol) supplemented with a protease inhibitor cocktail (Selleck) for 30 min with gentle agitation. ..

    Protease Inhibitor:

    Article Title: Cryo-EM structures of human P2X2/3 heteromer channel reveal the structural basis of ligand selectivity
    Article Snippet: .. For P2X2 or P2X3 protein purification, 500-ml cell pellets were resuspended in 25 ml of hypotonic buffer (25 mM Hepes, pH 7.5, 150 mM NaCl, 5 mM CaCl 2 , 2 mM TCEP, and 10% glycerol) supplemented with a protease inhibitor cocktail (Selleck) for 30 min with gentle agitation. ..

    Article Title: Cryo-EM structures of human P2X2/3 heteromer channel reveal the structural basis of ligand selectivity.
    Article Snippet: .. For P2X2 or P2X3 protein purification, 500- ml cell pellets were resuspended in 25 ml of hypotonic buffer (25 mM Hepes, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 2 mM TCEP, and 10% glycerol) supplemented with a protease inhibitor cocktail (Selleck) for 30 min with gentle agitation. ..

    Gentle:

    Article Title: Cryo-EM structures of human P2X2/3 heteromer channel reveal the structural basis of ligand selectivity
    Article Snippet: .. For P2X2 or P2X3 protein purification, 500-ml cell pellets were resuspended in 25 ml of hypotonic buffer (25 mM Hepes, pH 7.5, 150 mM NaCl, 5 mM CaCl 2 , 2 mM TCEP, and 10% glycerol) supplemented with a protease inhibitor cocktail (Selleck) for 30 min with gentle agitation. ..

    Article Title: Cryo-EM structures of human P2X2/3 heteromer channel reveal the structural basis of ligand selectivity.
    Article Snippet: .. For P2X2 or P2X3 protein purification, 500- ml cell pellets were resuspended in 25 ml of hypotonic buffer (25 mM Hepes, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 2 mM TCEP, and 10% glycerol) supplemented with a protease inhibitor cocktail (Selleck) for 30 min with gentle agitation. ..



    Similar Products

    93
    Selleck Chemicals buffer 2 mm tcep
    Buffer 2 Mm Tcep, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/buffer+2+mm+tcep/Glycerol/pm41931608-290-36-41
    Average 93 stars, based on 1 article reviews
    buffer 2 mm tcep - by Bioz Stars, 2026-09
    93/100 stars
      Buy from Supplier

    90
    Fisher Scientific coacervate buffer (20 mm hepes, 100 mm kcl, ph 7.5, freshly prepared with 100 µm tris(2-carboxyethyl)phosphine (tcep)
    a Schematic overview of affinity-based uptake of a client peptide of the 14-3-3 hub protein in a specific <t>coacervate</t> population. Two isoforms of His-tagged 14-3-3 are anchored in separate terpolymer-stabilized coacervates by means of their interaction with Ni-NTA-amylose in the coacervates. b Schematic overview of inter-coacervate signaling based on 14-3-3 interactions. PKA phosphorylates a phosphorylation-dependent and coacervate-anchored client protein (competitor, gray), which displaces an initially bound, but mobile, client protein (moderate binder, green) and facilitates the release of the mobile client protein into bulk solution. Subsequently, the green client protein is taken up into the receiver population of coacervates. 14-3-3, PKA, and the phosphorylation-dependent client protein are immobilized in the coacervates by means of interactions between their His-tag and Ni-NTA-amyloses.
    Coacervate Buffer (20 Mm Hepes, 100 Mm Kcl, Ph 7.5, Freshly Prepared With 100 µm Tris(2 Carboxyethyl)phosphine (Tcep), supplied by Fisher Scientific, 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/product/buffer+2+mm+tcep/tris+2+carboxyethyl+phosphine/pmc11213860-174-7-36
    Average 90 stars, based on 1 article reviews
    coacervate buffer (20 mm hepes, 100 mm kcl, ph 7.5, freshly prepared with 100 µm tris(2-carboxyethyl)phosphine (tcep) - by Bioz Stars, 2026-09
    90/100 stars
      Buy from Supplier

    99
    Thermo Fisher buffer agp 0 1 mm nah 2 po 4 and 50 mm tcep rbd 3 m urea and 250 mm tcep ph 2 8
    a Schematic overview of affinity-based uptake of a client peptide of the 14-3-3 hub protein in a specific <t>coacervate</t> population. Two isoforms of His-tagged 14-3-3 are anchored in separate terpolymer-stabilized coacervates by means of their interaction with Ni-NTA-amylose in the coacervates. b Schematic overview of inter-coacervate signaling based on 14-3-3 interactions. PKA phosphorylates a phosphorylation-dependent and coacervate-anchored client protein (competitor, gray), which displaces an initially bound, but mobile, client protein (moderate binder, green) and facilitates the release of the mobile client protein into bulk solution. Subsequently, the green client protein is taken up into the receiver population of coacervates. 14-3-3, PKA, and the phosphorylation-dependent client protein are immobilized in the coacervates by means of interactions between their His-tag and Ni-NTA-amyloses.
    Buffer Agp 0 1 Mm Nah 2 Po 4 And 50 Mm Tcep Rbd 3 M Urea And 250 Mm Tcep Ph 2 8, 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/product/buffer+2+mm+tcep/Urea/pmc10830291-85-43-86
    Average 99 stars, based on 1 article reviews
    buffer agp 0 1 mm nah 2 po 4 and 50 mm tcep rbd 3 m urea and 250 mm tcep ph 2 8 - by Bioz Stars, 2026-09
    99/100 stars
      Buy from Supplier

    90
    Merck KGaA benzonase elution buffer (20 mm tris ph 8.5, 2 mm mgcl 2 , 0.05% n-laroylsarcosine (nls), 0.5 mm tcep)
    SND1 interacts with RTC components involved in viral RNA biogenesis (A) Strategy to globally identify SND1 protein-protein interactome changes upon SARS-CoV-2 infection. (B) Fold change correlation plot displaying SND1 interacting proteins enriched over IgG control in SARS-CoV-2 infected (SCoV-2, y axis) and uninfected (mock, x axis) A549 ACE2 cells (n = 2 independent experiments). Candidates with a fold change > 1.5 or < 0.66 and FDR < 0.2 in infected relative to uninfected cells are displayed. Proteins with a substantial SND1 interaction change (absolute log 2 fold change > 1, FDR < 0.05) are highlighted in red and blue. (C) Computational slices of representative electron tomograms of SND1 knockout (KO) and control (CTRL) cells infected with SARS-CoV-2. Zoom in to region containing DMVs is shown at different magnifications. Left: scale bars, 1 μm; center: scale bars, 250 nm; right: scale bars, 100 nm. (D) Quantification of cross-sectional DMV area in SND1 KO and CTRL cells. Box: 25 th and 75 th percentiles. Whiskers: minimum to maximum. Median indicated by line. p value determined by t test, ∗∗ p < 0.01. (E) Proximity ligation assay for SND1 and NSP9. Mock-infected cells are compared to SARS-CoV-2 infected cells. Scale bars, 30 μm. (F) CoIP western blot analysis for epitope-tagged SND1 and NSP9 proteins expressed in uninfected HEK293T cells with and without nuclease <t>(benzonase)</t> treatment. SND1-FLAG served as bait. Input lysates are shown on the left. (G) As in (F), but using NSP9-V5 as bait. See also <xref ref-type=Figure S5 and Table S4 . " width="250" height="auto" />
    Benzonase Elution Buffer (20 Mm Tris Ph 8.5, 2 Mm Mgcl 2 , 0.05% N Laroylsarcosine (Nls), 0.5 Mm Tcep), 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/product/buffer+2+mm+tcep/mgcl2/pmc10617981-566-7-35
    Average 90 stars, based on 1 article reviews
    benzonase elution buffer (20 mm tris ph 8.5, 2 mm mgcl 2 , 0.05% n-laroylsarcosine (nls), 0.5 mm tcep) - by Bioz Stars, 2026-09
    90/100 stars
      Buy from Supplier

    90
    Thermo Fisher pic-dialysis buffer (20 mm hepes ph 7.5, 75 mm kcl, 1% glycerol, 2 mm mgcl2, 1 mm tcep)
    SND1 interacts with RTC components involved in viral RNA biogenesis (A) Strategy to globally identify SND1 protein-protein interactome changes upon SARS-CoV-2 infection. (B) Fold change correlation plot displaying SND1 interacting proteins enriched over IgG control in SARS-CoV-2 infected (SCoV-2, y axis) and uninfected (mock, x axis) A549 ACE2 cells (n = 2 independent experiments). Candidates with a fold change > 1.5 or < 0.66 and FDR < 0.2 in infected relative to uninfected cells are displayed. Proteins with a substantial SND1 interaction change (absolute log 2 fold change > 1, FDR < 0.05) are highlighted in red and blue. (C) Computational slices of representative electron tomograms of SND1 knockout (KO) and control (CTRL) cells infected with SARS-CoV-2. Zoom in to region containing DMVs is shown at different magnifications. Left: scale bars, 1 μm; center: scale bars, 250 nm; right: scale bars, 100 nm. (D) Quantification of cross-sectional DMV area in SND1 KO and CTRL cells. Box: 25 th and 75 th percentiles. Whiskers: minimum to maximum. Median indicated by line. p value determined by t test, ∗∗ p < 0.01. (E) Proximity ligation assay for SND1 and NSP9. Mock-infected cells are compared to SARS-CoV-2 infected cells. Scale bars, 30 μm. (F) CoIP western blot analysis for epitope-tagged SND1 and NSP9 proteins expressed in uninfected HEK293T cells with and without nuclease <t>(benzonase)</t> treatment. SND1-FLAG served as bait. Input lysates are shown on the left. (G) As in (F), but using NSP9-V5 as bait. See also <xref ref-type=Figure S5 and Table S4 . " width="250" height="auto" />
    Pic Dialysis Buffer (20 Mm Hepes Ph 7.5, 75 Mm Kcl, 1% Glycerol, 2 Mm Mgcl2, 1 Mm Tcep), supplied by Thermo Fisher, 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/product/buffer+2+mm+tcep/10__1016_slash_j__scp__2023__101100-174-14-42
    Average 90 stars, based on 1 article reviews
    pic-dialysis buffer (20 mm hepes ph 7.5, 75 mm kcl, 1% glycerol, 2 mm mgcl2, 1 mm tcep) - by Bioz Stars, 2026-09
    90/100 stars
      Buy from Supplier

    90
    Millipore 1.2 ml rna lysis and stabilization buffer [4 m licl, 5% triton-x100, 5% dgme, 10 mm edta, 50 mm tcep, 1% na 2 wo 4 , 100 mm hepes at ph 8.8]
    SND1 interacts with RTC components involved in viral RNA biogenesis (A) Strategy to globally identify SND1 protein-protein interactome changes upon SARS-CoV-2 infection. (B) Fold change correlation plot displaying SND1 interacting proteins enriched over IgG control in SARS-CoV-2 infected (SCoV-2, y axis) and uninfected (mock, x axis) A549 ACE2 cells (n = 2 independent experiments). Candidates with a fold change > 1.5 or < 0.66 and FDR < 0.2 in infected relative to uninfected cells are displayed. Proteins with a substantial SND1 interaction change (absolute log 2 fold change > 1, FDR < 0.05) are highlighted in red and blue. (C) Computational slices of representative electron tomograms of SND1 knockout (KO) and control (CTRL) cells infected with SARS-CoV-2. Zoom in to region containing DMVs is shown at different magnifications. Left: scale bars, 1 μm; center: scale bars, 250 nm; right: scale bars, 100 nm. (D) Quantification of cross-sectional DMV area in SND1 KO and CTRL cells. Box: 25 th and 75 th percentiles. Whiskers: minimum to maximum. Median indicated by line. p value determined by t test, ∗∗ p < 0.01. (E) Proximity ligation assay for SND1 and NSP9. Mock-infected cells are compared to SARS-CoV-2 infected cells. Scale bars, 30 μm. (F) CoIP western blot analysis for epitope-tagged SND1 and NSP9 proteins expressed in uninfected HEK293T cells with and without nuclease <t>(benzonase)</t> treatment. SND1-FLAG served as bait. Input lysates are shown on the left. (G) As in (F), but using NSP9-V5 as bait. See also <xref ref-type=Figure S5 and Table S4 . " width="250" height="auto" />
    1.2 Ml Rna Lysis And Stabilization Buffer [4 M Licl, 5% Triton X100, 5% Dgme, 10 Mm Edta, 50 Mm Tcep, 1% Na 2 Wo 4 , 100 Mm Hepes At Ph 8.8], 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/product/buffer+2+mm+tcep/triton+x+100/pmc03561265-169-11-43
    Average 90 stars, based on 1 article reviews
    1.2 ml rna lysis and stabilization buffer [4 m licl, 5% triton-x100, 5% dgme, 10 mm edta, 50 mm tcep, 1% na 2 wo 4 , 100 mm hepes at ph 8.8] - by Bioz Stars, 2026-09
    90/100 stars
      Buy from Supplier

    98
    Thermo Fisher buffer 100 mm nacl 1 mm tcep
    SND1 interacts with RTC components involved in viral RNA biogenesis (A) Strategy to globally identify SND1 protein-protein interactome changes upon SARS-CoV-2 infection. (B) Fold change correlation plot displaying SND1 interacting proteins enriched over IgG control in SARS-CoV-2 infected (SCoV-2, y axis) and uninfected (mock, x axis) A549 ACE2 cells (n = 2 independent experiments). Candidates with a fold change > 1.5 or < 0.66 and FDR < 0.2 in infected relative to uninfected cells are displayed. Proteins with a substantial SND1 interaction change (absolute log 2 fold change > 1, FDR < 0.05) are highlighted in red and blue. (C) Computational slices of representative electron tomograms of SND1 knockout (KO) and control (CTRL) cells infected with SARS-CoV-2. Zoom in to region containing DMVs is shown at different magnifications. Left: scale bars, 1 μm; center: scale bars, 250 nm; right: scale bars, 100 nm. (D) Quantification of cross-sectional DMV area in SND1 KO and CTRL cells. Box: 25 th and 75 th percentiles. Whiskers: minimum to maximum. Median indicated by line. p value determined by t test, ∗∗ p < 0.01. (E) Proximity ligation assay for SND1 and NSP9. Mock-infected cells are compared to SARS-CoV-2 infected cells. Scale bars, 30 μm. (F) CoIP western blot analysis for epitope-tagged SND1 and NSP9 proteins expressed in uninfected HEK293T cells with and without nuclease <t>(benzonase)</t> treatment. SND1-FLAG served as bait. Input lysates are shown on the left. (G) As in (F), but using NSP9-V5 as bait. See also <xref ref-type=Figure S5 and Table S4 . " width="250" height="auto" />
    Buffer 100 Mm Nacl 1 Mm Tcep, supplied by Thermo Fisher, 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/product/buffer+2+mm+tcep/Tris(2-carboxyethyl)phosphine+hydrochloride/pmc08387046-363-16-41
    Average 98 stars, based on 1 article reviews
    buffer 100 mm nacl 1 mm tcep - by Bioz Stars, 2026-09
    98/100 stars
      Buy from Supplier

    Image Search Results


    a Schematic overview of affinity-based uptake of a client peptide of the 14-3-3 hub protein in a specific coacervate population. Two isoforms of His-tagged 14-3-3 are anchored in separate terpolymer-stabilized coacervates by means of their interaction with Ni-NTA-amylose in the coacervates. b Schematic overview of inter-coacervate signaling based on 14-3-3 interactions. PKA phosphorylates a phosphorylation-dependent and coacervate-anchored client protein (competitor, gray), which displaces an initially bound, but mobile, client protein (moderate binder, green) and facilitates the release of the mobile client protein into bulk solution. Subsequently, the green client protein is taken up into the receiver population of coacervates. 14-3-3, PKA, and the phosphorylation-dependent client protein are immobilized in the coacervates by means of interactions between their His-tag and Ni-NTA-amyloses.

    Journal: Communications Chemistry

    Article Title: Competitive protein recruitment in artificial cells

    doi: 10.1038/s42004-024-01229-9

    Figure Lengend Snippet: a Schematic overview of affinity-based uptake of a client peptide of the 14-3-3 hub protein in a specific coacervate population. Two isoforms of His-tagged 14-3-3 are anchored in separate terpolymer-stabilized coacervates by means of their interaction with Ni-NTA-amylose in the coacervates. b Schematic overview of inter-coacervate signaling based on 14-3-3 interactions. PKA phosphorylates a phosphorylation-dependent and coacervate-anchored client protein (competitor, gray), which displaces an initially bound, but mobile, client protein (moderate binder, green) and facilitates the release of the mobile client protein into bulk solution. Subsequently, the green client protein is taken up into the receiver population of coacervates. 14-3-3, PKA, and the phosphorylation-dependent client protein are immobilized in the coacervates by means of interactions between their His-tag and Ni-NTA-amyloses.

    Article Snippet: The combined fractions were extensively dialyzed against coacervate buffer (20 mM HEPES, 100 mM KCl, pH 7.5, freshly prepared with 100 µM tris(2-carboxyethyl)phosphine (TCEP) using membrane tubing with a molecular weight cut-off (MWCO) of 12–14 kDa (Fisher Scientific).

    Techniques:

    a Schematic overview of binding of the FITC-c-Raf pS peptide to 14-3-3 isoforms and uptake of FITC-c-Raf pS in individual populations of coacervates containing either the weak 14-3-3σ isoform or the strong 14-3-3γ isoform. b Confocal micrographs showing the uptake of FITC-c-Raf pS in individual populations in the absence or presence of different 14-3-3 isoforms. Scale bar: 25 µm. Uncropped images are available in Supplementary Fig . c Schematic overview of competitive uptake of FITC-c-Raf pS into the coacervates containing two 14-3-3 isoforms with different affinity. d Confocal micrograph of competitive FITC-c-Raf pS uptake in a mixed coacervate sample containing coacervates loaded with either 14-3-3σ or with 14-3-3γ. Scale bar: 25 µm. Uncropped images are available in Supplementary Fig. . e , f Quantification of the FITC-c-Raf pS signal from micrographs of individual populations ( e ) or mixed ( f ) coacervate populations containing different 14-3-3 isoforms. Statistical differences were analyzed by nonparametric Dunn’s test with correction for multiple comparisons, with N ≥ 21 coacervates across multiple imaging positions in the same sample. P values are shown above the comparison. Dashed lines represent the median and dotted lines represent the upper and lower quartiles. ns: no statistical difference. The fold difference is given as the fold difference between the means.

    Journal: Communications Chemistry

    Article Title: Competitive protein recruitment in artificial cells

    doi: 10.1038/s42004-024-01229-9

    Figure Lengend Snippet: a Schematic overview of binding of the FITC-c-Raf pS peptide to 14-3-3 isoforms and uptake of FITC-c-Raf pS in individual populations of coacervates containing either the weak 14-3-3σ isoform or the strong 14-3-3γ isoform. b Confocal micrographs showing the uptake of FITC-c-Raf pS in individual populations in the absence or presence of different 14-3-3 isoforms. Scale bar: 25 µm. Uncropped images are available in Supplementary Fig . c Schematic overview of competitive uptake of FITC-c-Raf pS into the coacervates containing two 14-3-3 isoforms with different affinity. d Confocal micrograph of competitive FITC-c-Raf pS uptake in a mixed coacervate sample containing coacervates loaded with either 14-3-3σ or with 14-3-3γ. Scale bar: 25 µm. Uncropped images are available in Supplementary Fig. . e , f Quantification of the FITC-c-Raf pS signal from micrographs of individual populations ( e ) or mixed ( f ) coacervate populations containing different 14-3-3 isoforms. Statistical differences were analyzed by nonparametric Dunn’s test with correction for multiple comparisons, with N ≥ 21 coacervates across multiple imaging positions in the same sample. P values are shown above the comparison. Dashed lines represent the median and dotted lines represent the upper and lower quartiles. ns: no statistical difference. The fold difference is given as the fold difference between the means.

    Article Snippet: The combined fractions were extensively dialyzed against coacervate buffer (20 mM HEPES, 100 mM KCl, pH 7.5, freshly prepared with 100 µM tris(2-carboxyethyl)phosphine (TCEP) using membrane tubing with a molecular weight cut-off (MWCO) of 12–14 kDa (Fisher Scientific).

    Techniques: Binding Assay, Imaging, Comparison

    SND1 interacts with RTC components involved in viral RNA biogenesis (A) Strategy to globally identify SND1 protein-protein interactome changes upon SARS-CoV-2 infection. (B) Fold change correlation plot displaying SND1 interacting proteins enriched over IgG control in SARS-CoV-2 infected (SCoV-2, y axis) and uninfected (mock, x axis) A549 ACE2 cells (n = 2 independent experiments). Candidates with a fold change > 1.5 or < 0.66 and FDR < 0.2 in infected relative to uninfected cells are displayed. Proteins with a substantial SND1 interaction change (absolute log 2 fold change > 1, FDR < 0.05) are highlighted in red and blue. (C) Computational slices of representative electron tomograms of SND1 knockout (KO) and control (CTRL) cells infected with SARS-CoV-2. Zoom in to region containing DMVs is shown at different magnifications. Left: scale bars, 1 μm; center: scale bars, 250 nm; right: scale bars, 100 nm. (D) Quantification of cross-sectional DMV area in SND1 KO and CTRL cells. Box: 25 th and 75 th percentiles. Whiskers: minimum to maximum. Median indicated by line. p value determined by t test, ∗∗ p < 0.01. (E) Proximity ligation assay for SND1 and NSP9. Mock-infected cells are compared to SARS-CoV-2 infected cells. Scale bars, 30 μm. (F) CoIP western blot analysis for epitope-tagged SND1 and NSP9 proteins expressed in uninfected HEK293T cells with and without nuclease (benzonase) treatment. SND1-FLAG served as bait. Input lysates are shown on the left. (G) As in (F), but using NSP9-V5 as bait. See also <xref ref-type=Figure S5 and Table S4 . " width="100%" height="100%">

    Journal: Cell

    Article Title: SND1 binds SARS-CoV-2 negative-sense RNA and promotes viral RNA synthesis through NSP9

    doi: 10.1016/j.cell.2023.09.002

    Figure Lengend Snippet: SND1 interacts with RTC components involved in viral RNA biogenesis (A) Strategy to globally identify SND1 protein-protein interactome changes upon SARS-CoV-2 infection. (B) Fold change correlation plot displaying SND1 interacting proteins enriched over IgG control in SARS-CoV-2 infected (SCoV-2, y axis) and uninfected (mock, x axis) A549 ACE2 cells (n = 2 independent experiments). Candidates with a fold change > 1.5 or < 0.66 and FDR < 0.2 in infected relative to uninfected cells are displayed. Proteins with a substantial SND1 interaction change (absolute log 2 fold change > 1, FDR < 0.05) are highlighted in red and blue. (C) Computational slices of representative electron tomograms of SND1 knockout (KO) and control (CTRL) cells infected with SARS-CoV-2. Zoom in to region containing DMVs is shown at different magnifications. Left: scale bars, 1 μm; center: scale bars, 250 nm; right: scale bars, 100 nm. (D) Quantification of cross-sectional DMV area in SND1 KO and CTRL cells. Box: 25 th and 75 th percentiles. Whiskers: minimum to maximum. Median indicated by line. p value determined by t test, ∗∗ p < 0.01. (E) Proximity ligation assay for SND1 and NSP9. Mock-infected cells are compared to SARS-CoV-2 infected cells. Scale bars, 30 μm. (F) CoIP western blot analysis for epitope-tagged SND1 and NSP9 proteins expressed in uninfected HEK293T cells with and without nuclease (benzonase) treatment. SND1-FLAG served as bait. Input lysates are shown on the left. (G) As in (F), but using NSP9-V5 as bait. See also Figure S5 and Table S4 .

    Article Snippet: To elute proteins, beads were rinsed with benzonase elution buffer (20 mM Tris pH 8.5, 2 mM MgCl 2 , 0.05% N-laroylsarcosine (NLS), 0.5 mM TCEP) while magnetically separated and incubated with 500 U benzonase (Merck Millipore, 71206-3) and 44 U RNase I (Thermo Fisher Scientific, AM2239) in benzonase elution buffer at 37°C and 1000 rpm overnight.

    Techniques: Infection, Knock-Out, Proximity Ligation Assay, Western Blot

    Journal: Cell

    Article Title: SND1 binds SARS-CoV-2 negative-sense RNA and promotes viral RNA synthesis through NSP9

    doi: 10.1016/j.cell.2023.09.002

    Figure Lengend Snippet:

    Article Snippet: To elute proteins, beads were rinsed with benzonase elution buffer (20 mM Tris pH 8.5, 2 mM MgCl 2 , 0.05% N-laroylsarcosine (NLS), 0.5 mM TCEP) while magnetically separated and incubated with 500 U benzonase (Merck Millipore, 71206-3) and 44 U RNase I (Thermo Fisher Scientific, AM2239) in benzonase elution buffer at 37°C and 1000 rpm overnight.

    Techniques: Virus, Recombinant, Concentration Assay, Protease Inhibitor, Magnetic Beads, Transfection, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Cell Viability Assay, In Situ, Immunohistochemistry, Sequencing, Plasmid Preparation, CRISPR, Software