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  • 99
    Thermo Fisher tbst
    <t>CPH1</t> migrates differently when overexpressed in E. coli or C. reinhardtii . A, E. coli cells transformed with pCPH101, which contains CPH1 cDNA, were induced with isopropylthio- β -galactoside for 2.5 h and cell extracts were analyzed using SDS-PAGE and stained with Coomassie Brilliant Blue. B, Western blot of E. coli pCPH101 extract before and after induction (left) and C. reinhardtii OxG extracts before and after heat shock (right). Western blots were performed using antibodies directed against CPH1 protein as described except <t>TBST</t> plus 0.3% (v/v) Triton X-100 was used.
    Tbst, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 38420 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore tbst
    <t>CPH1</t> migrates differently when overexpressed in E. coli or C. reinhardtii . A, E. coli cells transformed with pCPH101, which contains CPH1 cDNA, were induced with isopropylthio- β -galactoside for 2.5 h and cell extracts were analyzed using SDS-PAGE and stained with Coomassie Brilliant Blue. B, Western blot of E. coli pCPH101 extract before and after induction (left) and C. reinhardtii OxG extracts before and after heat shock (right). Western blots were performed using antibodies directed against CPH1 protein as described except <t>TBST</t> plus 0.3% (v/v) Triton X-100 was used.
    Tbst, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 27500 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tbst - by Bioz Stars, 2020-09
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    99
    Abcam tween 20 tbst
    <t>CPH1</t> migrates differently when overexpressed in E. coli or C. reinhardtii . A, E. coli cells transformed with pCPH101, which contains CPH1 cDNA, were induced with isopropylthio- β -galactoside for 2.5 h and cell extracts were analyzed using SDS-PAGE and stained with Coomassie Brilliant Blue. B, Western blot of E. coli pCPH101 extract before and after induction (left) and C. reinhardtii OxG extracts before and after heat shock (right). Western blots were performed using antibodies directed against CPH1 protein as described except <t>TBST</t> plus 0.3% (v/v) Triton X-100 was used.
    Tween 20 Tbst, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 4426 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Cell Signaling Technology Inc tween 20 tbst
    <t>CPH1</t> migrates differently when overexpressed in E. coli or C. reinhardtii . A, E. coli cells transformed with pCPH101, which contains CPH1 cDNA, were induced with isopropylthio- β -galactoside for 2.5 h and cell extracts were analyzed using SDS-PAGE and stained with Coomassie Brilliant Blue. B, Western blot of E. coli pCPH101 extract before and after induction (left) and C. reinhardtii OxG extracts before and after heat shock (right). Western blots were performed using antibodies directed against CPH1 protein as described except <t>TBST</t> plus 0.3% (v/v) Triton X-100 was used.
    Tween 20 Tbst, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 3703 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Santa Cruz Biotechnology tween 20 tbst
    <t>CPH1</t> migrates differently when overexpressed in E. coli or C. reinhardtii . A, E. coli cells transformed with pCPH101, which contains CPH1 cDNA, were induced with isopropylthio- β -galactoside for 2.5 h and cell extracts were analyzed using SDS-PAGE and stained with Coomassie Brilliant Blue. B, Western blot of E. coli pCPH101 extract before and after induction (left) and C. reinhardtii OxG extracts before and after heat shock (right). Western blots were performed using antibodies directed against CPH1 protein as described except <t>TBST</t> plus 0.3% (v/v) Triton X-100 was used.
    Tween 20 Tbst, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 4423 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore tween 20 tbst
    <t>CPH1</t> migrates differently when overexpressed in E. coli or C. reinhardtii . A, E. coli cells transformed with pCPH101, which contains CPH1 cDNA, were induced with isopropylthio- β -galactoside for 2.5 h and cell extracts were analyzed using SDS-PAGE and stained with Coomassie Brilliant Blue. B, Western blot of E. coli pCPH101 extract before and after induction (left) and C. reinhardtii OxG extracts before and after heat shock (right). Western blots were performed using antibodies directed against CPH1 protein as described except <t>TBST</t> plus 0.3% (v/v) Triton X-100 was used.
    Tween 20 Tbst, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2926 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Cell Signaling Technology Inc tbst
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Tbst, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 19645 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Bio-Rad tbst
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Tbst, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 10639 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Santa Cruz Biotechnology tbst
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Tbst, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 97/100, based on 24731 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tbst  (Abcam)
    94
    Abcam tbst
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Tbst, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 15271 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc tris buffered saline tween 20
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Tris Buffered Saline Tween 20, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 512 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore tbst buffer
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Tbst Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2242 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Bio-Rad tween 20 tbst
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Tween 20 Tbst, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 607 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam tween tbst
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Tween Tbst, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 509 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore bsa tbst
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Bsa Tbst, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 592 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Agilent technologies tbst
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Tbst, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 5326 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Santa Cruz Biotechnology tween tbst
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Tween Tbst, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 543 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cell Signaling Technology Inc tris buffered saline tween 20 tbst
    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted <t>1:3000</t> (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in <t>TBST.</t> The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p
    Tris Buffered Saline Tween 20 Tbst, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 421 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    CPH1 migrates differently when overexpressed in E. coli or C. reinhardtii . A, E. coli cells transformed with pCPH101, which contains CPH1 cDNA, were induced with isopropylthio- β -galactoside for 2.5 h and cell extracts were analyzed using SDS-PAGE and stained with Coomassie Brilliant Blue. B, Western blot of E. coli pCPH101 extract before and after induction (left) and C. reinhardtii OxG extracts before and after heat shock (right). Western blots were performed using antibodies directed against CPH1 protein as described except TBST plus 0.3% (v/v) Triton X-100 was used.

    Journal: Plant Physiology

    Article Title: The CPH1 Gene of Chlamydomonas reinhardtii Encodes Two Forms of Cryptochrome Whose Levels Are Controlled by Light-Induced Proteolysis 1 Encodes Two Forms of Cryptochrome Whose Levels Are Controlled by Light-Induced Proteolysis 1 [w]

    doi: 10.1104/pp.103.031930

    Figure Lengend Snippet: CPH1 migrates differently when overexpressed in E. coli or C. reinhardtii . A, E. coli cells transformed with pCPH101, which contains CPH1 cDNA, were induced with isopropylthio- β -galactoside for 2.5 h and cell extracts were analyzed using SDS-PAGE and stained with Coomassie Brilliant Blue. B, Western blot of E. coli pCPH101 extract before and after induction (left) and C. reinhardtii OxG extracts before and after heat shock (right). Western blots were performed using antibodies directed against CPH1 protein as described except TBST plus 0.3% (v/v) Triton X-100 was used.

    Article Snippet: Primary antibodies against CPH1 were diluted to 1:1,000 in 1% (w/v) bovine serum albumin in TBST and incubated with the membrane for 1 h. M2 monoclonal antibodies against FLAG (Invitrogen, Carlsbad, CA) were diluted 1:1000 in 1% bovine serum albumin, 2.5% NFDM in TBST, and incubated with the membrane for 1 h. Regardless of the primary antibody used, membranes were subsequently washed for 30 min in TBST or TBST plus 0.3% (v/v) Triton X-100.

    Techniques: Transformation Assay, SDS Page, Staining, Western Blot

    Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted 1:3000 (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in TBST. The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p

    Journal: Apoptosis

    Article Title: Accumulation of tissue factor in endothelial cells promotes cellular apoptosis through over-activation of Src1 and involves β1-integrin signalling

    doi: 10.1007/s10495-019-01576-2

    Figure Lengend Snippet: Analysis of Src phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP, b pCMV6-Ac-TF Ala253 -tGFP, c pCMV6-Ac-TF-tGFP plasmids, along with d an untransfected set of cells. The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed in Laemmeli’s buffer containing a protease inhibitor cocktail and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Src1 antibody, diluted 1:3000 (v/v) or a rabbit monoclonal anti-human phospho-Tyr416-Src family antibody (D49G4), diluted 1:4000 (v/v) in TBST. The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Src were calculated using the ImageJ program. (n = 6; * = p

    Article Snippet: Total and phosphorylated FAK were detected using a polyclonal rabbit anti-human FAK diluted 1:3000 (v/v) in TBST, and a monoclonal rabbit anti-human phospho-Tyr397-FAK (D20B1) diluted 1:3000 (v/v) in TBST, respectively (Cell Signalling Technologies).

    Techniques: Western Blot, Transfection, Incubation, Expressing, Recombinant, Protease Inhibitor, SDS Page

    Analysis of Rac phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP or b pCMV6-Ac-TF Ala253 -tGFP (additional samples shown in Supplementary Fig. 2). The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were then activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Rac1/2/3 antibody, diluted 1:3000 (v/v) and a rabbit monoclonal anti-human phospho-Ser71-Rac1 antibody, diluted 1:3000 (v/v) diluted in TBST. The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Rac were calculated using the ImageJ program. (n = 6; * = p

    Journal: Apoptosis

    Article Title: Accumulation of tissue factor in endothelial cells promotes cellular apoptosis through over-activation of Src1 and involves β1-integrin signalling

    doi: 10.1007/s10495-019-01576-2

    Figure Lengend Snippet: Analysis of Rac phosphorylation by western blot. Human coronary artery endothelial cells (2 × 10 5 ) were seeded out into 12-well plates and transfected with 0.5 µg of a pCMV6-Ac-tGFP or b pCMV6-Ac-TF Ala253 -tGFP (additional samples shown in Supplementary Fig. 2). The cells were incubated for 48 h to permit the expression of the recombinant proteins. Sets of cells were then activated with PAR2-AP (20 µM) and incubated for up to 120 min. The cells were then lysed and separated by 12% (w/v) SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were probed with a polyclonal rabbit anti-human Rac1/2/3 antibody, diluted 1:3000 (v/v) and a rabbit monoclonal anti-human phospho-Ser71-Rac1 antibody, diluted 1:3000 (v/v) diluted in TBST. The membranes were then washed and probed with a goat anti-rabbit alkaline phosphatase-conjugated antibody diluted 1:1000 (v/v) and then visualised using the Western Blue stabilised alkaline phosphatase-substrate and recorded. All quantifications were normalised against GAPDH which was detected using a polyclonal goat anti-GAPDH antibody diluted 1:5000 (v/v) and then detected using an alkaline phosphatase-conjugated donkey anti-goat-IgG antibody diluted 1:2000 (v/v). The ratios of phosphorylated to total Rac were calculated using the ImageJ program. (n = 6; * = p

    Article Snippet: Total and phosphorylated FAK were detected using a polyclonal rabbit anti-human FAK diluted 1:3000 (v/v) in TBST, and a monoclonal rabbit anti-human phospho-Tyr397-FAK (D20B1) diluted 1:3000 (v/v) in TBST, respectively (Cell Signalling Technologies).

    Techniques: Western Blot, Transfection, Incubation, Expressing, Recombinant, SDS Page