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Antibody against gapdh

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Antibody against GAPDH is a laboratory reagent used to detect the presence and quantify the levels of the GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) protein in biological samples. GAPDH is a commonly used housekeeping gene and its expression is often used as a reference point in various experimental techniques.

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Lab products found in correlation

Pvdf membrane, by Merck Group (3 mentions) Ripa lysis buffer, by Beyotime (3 mentions) Ecl chemiluminescence kit hrp, by Fdbio (2 mentions) Cleaved parp, by Cell Signaling Technology (2 mentions) Cleaved caspase 3, by Cell Signaling Technology (2 mentions) Supersignal west pico plus chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Enhanced bca protein assay kit, by Beyotime (1 mentions) N cadherin, by Cell Signaling Technology (1 mentions) Vimentin, by Cell Signaling Technology (1 mentions) Mcl 1, by Santa Cruz Biotechnology (1 mentions) Pchk2 t68, by Cell Signaling Technology (1 mentions) γh2ax, by Cell Signaling Technology (1 mentions) Caspase 8, by Cell Signaling Technology (1 mentions) Caspase 3, by Cell Signaling Technology (1 mentions) Ube2c, by Cell Signaling Technology (1 mentions) Mln7243, by MedChemExpress (1 mentions) P chk1 ser317, by Cell Signaling Technology (1 mentions) Cul4a, by Cell Signaling Technology (1 mentions) Mln4924, by MedChemExpress (1 mentions) Cleaved caspase 8, by Cell Signaling Technology (1 mentions)

5 protocols using antibody against gapdh

1

Western Blot Analysis of MAPK and STAT3

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After the different treatment, cells were lysed in RIPA lysis buffer (Beyotime, Shanghai, China). Then, the protein was separated by SDS‐PAGE, transferred to PVDF membranes (Millipore, Bedford, MA, USA) and probed with the indicated primary antibodies for overnight at 4℃. After incubated with the corresponding secondary antibodies for 1 hour, the densities of bands were examined by ECL Chemiluminescence Kit HRP (FDbio, Hangzhou, China). Antibodies against ERK, p‐ERK (Thr202/Thy204), p38, p‐p38(Thr180/Tyr182) were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against JNK, p‐JNK(Thr183) and p‐STAT3(Tyr705) were purchased from Diag Biotechnology (Hangzhou, China). The antibody against STAT3 was obtained from Affinity Biosciences (Cincinnati, OH, USA). The antibody against GAPDH was obtained from Beyotime (Shanghai, China).
Cui L., Yang G., Ye J., Yao Y., Lu G., Chen J., Fang L., Lu S, & Zhou J. (2020). Dioscin elicits anti‐tumour immunity by inhibiting macrophage M2 polarization via JNK and STAT3 pathways in lung cancer. Journal of Cellular and Molecular Medicine, 24(16), 9217-9230.
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2

Western Blot Protein Analysis

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Total cell lysates were prepared from either liver tissue or cultured cells by lysis using RIPA Lysis Buffer (Beyotime, China) containing 1% PMSF and homogenization using a vortex oscillator (Roche, USA). Protein concentrations were determined using an Enhanced BCA Protein Assay Kit (Beyotime, China). An equal volume of 5 X loading buffer was mixed with the samples, which were boiled for 10 minutes. After separation by 10% SDS-PAGE, the proteins were transferred to a 0.45 mm PVDF membrane and blocked for 2 hours with 5% non-fat dry milk at room temperature. Then, the membrane was incubated overnight at 4 °C with the appropriate primary antibody. After incubation with the secondary antibody for another hour, the membranes were developed with SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Thermo, USA). The following primary antibodies were used. Antibodies against ACOX1 (A8091) and CPT1A (A5307) were purchased from ABclonal, and an antibody against AHR (#4685) was purchased from Cell Signaling Technology (Danvers, MA, USA). The antibody against GAPDH (AG019) was purchased from Beyotime Biotechnology.
Han Q., Chen H., Wang L., An Y., Hu X., Zhao Y., Zhang H, & Zhang R. (2021). Systemic Deficiency of GHR in Pigs leads to Hepatic Steatosis via Negative Regulation of AHR Signaling. International Journal of Biological Sciences, 17(15), 4108-4121.
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3

Western Blot Analysis of Apoptosis and EMT Markers

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Cells and tissues were lysed in RIPA lysis buffer (Beyotime, Shanghai, China). Then, the protein was separated by SDS-PAGE, transferred to PVDF membranes (Millipore, Bedford, MA, USA) and probed with primary antibodies overnight at 4 ℃. After incubated with the corresponding secondary antibodies for 1 h, the densities of bands were detected by ECL Chemiluminescence Kit HRP (FDbio, Hangzhou, China). Antibodies against Bcl2, Bax, cleaved caspase-3, cleaved PARP, p38, p-p38 (Thr180/Tyr182), E-cadherin, N-cadherin and Vimentin were purchased from Cell Signalling Technology (Danvers, MA, USA). Antibodies against HSP27 and p-HSP27 (Ser15) were purchased from Diag Biotechnology (Hangzhou, China). The antibody against GAPDH was obtained from Beyotime (Shanghai, China).
Yao Y., Cui L., Ye J., Yang G., Lu G., Fang X., Zeng Z, & Zhou J. (2020). Dioscin facilitates ROS-induced apoptosis via the p38-MAPK/HSP27-mediated pathways in lung squamous cell carcinoma. International Journal of Biological Sciences, 16(15), 2883-2894.
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4

Assessing Protein Ubiquitination Dynamics

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MLN4924 and MLN7243 were purchased from MedChemExpress (Monmouth Junction, NJ, USA). Drugs were dissolved in dimethyl sulfoxide (DMSO), aliquoted, and stored at -20°C.
Primary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA) against the following: Ubc12 (#5641S); UBE2C (#14234S); Ubc9 (#4786S); Cul3 (#2759S); Cul4A (#2699S); NAE1/APPBP1 (#14321S); Cdt1 (#8064S); p27 (#3688S); p-Chk1-Ser317 (#2344S); Chk1 (#2360S); p-Chk2-T68 (#2661S); γH2AX (#2577L); Caspase-8 (#4790S); cleaved Caspase-8 (#9748L); caspase-3 (#9662S); cleaved caspase-3 (#9661S); caspase-7 (#9492T); cleaved PARP (#5625S); BAK (#12105S); Noxa (#14766S); Puma (#4976S); BAD (#9292S); BID (#2002S); BCL-XL (#2764S). Antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA) against the following: UBA3 (sc-377272); Cul1 (sc-17775); Wee1 (sc-5285); p21 (sc-271610); Chk2 (sc-9604); PARP1 (sc-7150); BAX (sc-493); BCL-2 (sc-492); MCL-1 (sc-819). Antibody against Cul4B (C9995) was from MilliporeSigma (Burlington, Massachusetts, US). Antibody against GAPDH was purchased from Beyotime Biotechnology (Shanghai, China). The secondary antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA).
Zhou L.N., Xiong C., Cheng Y.J., Song S.S., Bao X.B., Huan X.J., Wang T.Y., Zhang A., Miao Z.H, & He J.X. (2022). SOMCL-19-133, a novel, selective, and orally available inhibitor of NEDD8-activating enzyme (NAE) for cancer therapy. Neoplasia (New York, N.Y.), 32, 100823.
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5

NUF2 Protein Expression Analysis

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A lysis buffer (Beyotime Institute of Biotechnology, Nantong, China) was used to prepare total protein extracts from cell lines as well as tumor tissues. Then, protein concentrations were evaluated by a BCA kit (Beyotime Institute of Biotechnology, Nantong, China). An equal protein amount (40 μg per lane) was separated by SDS-polyacrylamide gel electrophoresis (PAGE) in 12% acrylamide gels and transferred onto polyvinylidine difluoride (PVDF) membranes (Millipore Corporation, Billerica, USA) which were blocked in 5% fat-free milk followed by overnight incubation at 4°C with the rabbit anti-NUF2 primary antibody (1 : 5000 dilution; Sangon Biotech, Shanghai, China). The secondary antibody was horseradish peroxidase- (HRP-) conjugated goat anti-rabbit antibody (1 : 2000; Beyotime Institute of Biotechnology). After stripping, the membrane was reprobed overnight at 4°C with antibody against GAPDH (1 : 2000; Beyotime Institute of Biotechnology), followed by incubation with secondary antibodies as above at room temperature (RT) for 2 h. An enhanced chemiluminescence system (ECL; Beyotime Institute of Biotechnology) was used for band visualization. The band intensities were quantified by densitometry.
Li X., Zhang L., Yi Z., Zhou J., Song W., Zhao P., Wu J., Song J, & Ni Q. (2022). NUF2 Is a Potential Immunological and Prognostic Marker for Non-Small-Cell Lung Cancer. Journal of Immunology Research, 2022, 1161931.
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