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Gst antibody sc 138

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The GST antibody (#sc-138) is a protein-specific antibody developed by Santa Cruz Biotechnology. It is designed to detect the glutathione S-transferase (GST) protein, a commonly used fusion tag in recombinant protein expression and purification.

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Lab products found in correlation

Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions) Flag f1804 antibody, by Merck Group (2 mentions) Nitrocellulose membrane, by GE Healthcare (2 mentions) Ecl detection reagent, by Cytiva (2 mentions) P 4e bp1 t37 46, by Cell Signaling Technology (2 mentions) β actin, by Cell Signaling Technology (2 mentions) Protease and phosphatase inhibitor, by Thermo Fisher Scientific (2 mentions) P 4ebp1 s65, by Cell Signaling Technology (2 mentions) P 4ebp1 t70, by Cell Signaling Technology (2 mentions) Raptor, by Cell Signaling Technology (2 mentions) Flag antibody, by Merck Group (1 mentions) Glutathione sepharose 4b, by GE Healthcare (1 mentions) Flag m2 agarose beads, by Merck Group (1 mentions)

Spelling variants of this product

Sc-138 (29 citations) GST (sc-138), (8 citations) GST antibody (6 citations)

4 protocols using gst antibody sc 138

1

Immunoblotting Analysis of Protein Phosphorylation

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Cells were washed with PBS once, disrupted on ice for thirty minutes in NP-40 (50 mM Tris [pH 7.4], 1% NP-40, 150 mmol/L NaCl, 40 mmol/L NaF) or RIPA lysis buffer (Thermo Scientific) supplemented with protease and phosphatase inhibitors (Pierce Chemical) and cleared by centrifugation. Protein concentration was determined with BCA reagent from Pierce. Equal amounts of protein (10 to 50 µg) in cell lysates were separated by SDS-PAGE, transferred to nitrocellulose membranes (GE healthcare), immunoblotted with specific primary and secondary antibodies and detected by chemiluminescence with the ECL detection reagents from Amersham Biosciences. Antibodies for P-AKT (S473) (#4060L), P-p70S6K (T389) (#9234L), P-S6 (S240/244) (#5364L) and P-S6 (S235/236) (#4858L), P-4EBP1 (T37/46) (#9459L), P-4EBP1 (S65) (#9451L), P-4EBP1 (T70) (#9455L), β-actin (#4970S), mTOR (#2972S) and Raptor (#2280S) were purchased from Cell Signaling Technology. The FLAG (#F1804) antibody was purchased from Sigma. The GST antibody (#sc-138) was from Santa Cruz.
Rodrik-Outmezguine V.S., Okaniwa M., Yao Z., Novotny C.J., McWhirter C., Banaji A., Won H., Wong W., Berger M., de Stanchina E., Barratt D.G., Cosulich S., Klinowska T., Rosen N, & Shokat K.M. (2016). Overcoming mTOR Resistance Mutations with a New Generation mTOR Inhibitor. Nature, 534(7606), 272-276.
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2

In Vitro LATS1 Kinase Assay

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GST fusion YAP proteins (2 μg) were used for the in vitro kinase assay. LATS1 kinase was immunoprecipitated (3477S, Cell Signaling Technology, 1:100 dilution for immunoprecipitation) from the indicated cell lysates and subjected to the kinase assay in the presence of cold ATP (500 μM) and GST-YAP fusion protein. The reaction mixture was incubated at 30 °C for 30 min, terminated with SDS loading buffer and subjected to SDS–PAGE. Phosphorylation of YAP at the S127 site was determined by YAP S127 phospho-antibody (4911S, Cell Signaling Technology, 1:1,000 dilution). GST antibody (sc-138, 1:1,000) was from Santa Cruz Biotechnology.
Yi J., Lu L., Yanger K., Wang W., Sohn B.H., Stanger B.Z., Zhang M., Martin J.F., Ajani J.A., Chen J., Lee J.S., Song S, & Johnson R.L. (2016). LATS1 and LATS2 regulate mouse liver progenitor cell proliferation and maturation through antagonism of the coactivators YAP and TAZ. Hepatology (Baltimore, Md.), 64(5), 1757-1772.
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3

Glutathione S-Transferase Pull-Down and Immunoprecipitation

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The glutathione S-transferase (GST) pull-down assays and immunoprecipitation experiments were performed as described before57 (link). Cells were mainly lysed in NP-40 buffer (0.4 M NaCl, 0.1% NP-40, 10 mM Tris-HCl, pH 8.0 and 1 mM EDTA) supplemented with protease inhibitors. Immunoprecipitation of the WASH complex and retromer was performed in MRB lysis buffer (20 mM HEPES pH 7.2, 50 mM potassium acetate, 1 mM EDTA, 200 mM D-sorbitol and 0.1% Triton X-100) supplemented with protease inhibitors. Equal amount of protein was used for immunoprecipiation and pull-down assays. FLAG M2 agarose beads (Sigma-Aldrich) were used for Flag-immunoprecipitation experiments, Glutathione Sepharose 4B (GE Healthcare Life Sciences) for GST pull-down assays. Flag-, Ha-, GST-tagged proteins were detected by Flag-antibody (F1804, Sigma, 1:2,000), Ha-antibody (clone HA-7, H6533, Sigma-Aldrich, 1:5,000) and GST-antibody (sc-138, Santa Cruz, 1:5,000), respectively.
Bartuzi P., Billadeau D.D., Favier R., Rong S., Dekker D., Fedoseienko A., Fieten H., Wijers M., Levels J.H., Huijkman N., Kloosterhuis N., van der Molen H., Brufau G., Groen A.K., Elliott A.M., Kuivenhoven J.A., Plecko B., Grangl G., McGaughran J., Horton J.D., Burstein E., Hofker M.H, & van de Sluis B. (2016). CCC- and WASH-mediated endosomal sorting of LDLR is required for normal clearance of circulating LDL. Nature Communications, 7, 10961.
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4

Immunoblotting Analysis of Protein Phosphorylation

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Cells were washed with PBS once, disrupted on ice for thirty minutes in NP-40 (50 mM Tris [pH 7.4], 1% NP-40, 150 mmol/L NaCl, 40 mmol/L NaF) or RIPA lysis buffer (Thermo Scientific) supplemented with protease and phosphatase inhibitors (Pierce Chemical) and cleared by centrifugation. Protein concentration was determined with BCA reagent from Pierce. Equal amounts of protein (10 to 50 µg) in cell lysates were separated by SDS-PAGE, transferred to nitrocellulose membranes (GE healthcare), immunoblotted with specific primary and secondary antibodies and detected by chemiluminescence with the ECL detection reagents from Amersham Biosciences. Antibodies for P-AKT (S473) (#4060L), P-p70S6K (T389) (#9234L), P-S6 (S240/244) (#5364L) and P-S6 (S235/236) (#4858L), P-4EBP1 (T37/46) (#9459L), P-4EBP1 (S65) (#9451L), P-4EBP1 (T70) (#9455L), β-actin (#4970S), mTOR (#2972S) and Raptor (#2280S) were purchased from Cell Signaling Technology. The FLAG (#F1804) antibody was purchased from Sigma. The GST antibody (#sc-138) was from Santa Cruz.
Rodrik-Outmezguine V.S., Okaniwa M., Yao Z., Novotny C.J., McWhirter C., Banaji A., Won H., Wong W., Berger M., de Stanchina E., Barratt D.G., Cosulich S., Klinowska T., Rosen N, & Shokat K.M. (2016). Overcoming mTOR Resistance Mutations with a New Generation mTOR Inhibitor. Nature, 534(7606), 272-276.
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