Epr6893

Manufactured by Abcam

EPR6893 is a monoclonal antibody that recognizes the human nuclear factor erythroid 2-related factor 2 (Nrf2) protein. Nrf2 is a transcription factor that plays a crucial role in the regulation of antioxidant response genes.

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Lab products found in correlation

3 protocols using epr6893

Western Blot Analysis of Cell Lysates

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Cells were lysed in protease inhibitor-supplemented RIPA buffer by rotation at 4°C for 30 minutes, followed by centrifugation at 14,000 g for 5 minutes at 4°C. Protein concentration was determined by BCA assay (Pierce Biotechnology) according to manufacturer’s instructions. Protein lysates (10 – 20 μg) were resolved by SDS-PAGE, transferred to PVDF (polyvinylidene difluoride) membranes, and probed with the indicated primary antibodies: α-tubulin (ab4074, Abcam), β-actin (AC-15, A3854, Sigma), BLNK (2B11, sc-8003, Santa Cruz), CAD (RB18384, AP11110c-ev, Abgent), CLPP (B-12, sc-271284, Santa Cruz), CTPS1 (C-13, sc-131474, Santa Cruz), G6PD (ab76598, Abcam), GAPDH (6C5, MAB374, Millipore Sigma), Hexokinase 2 (C-14, sc-6521, Santa Cruz), HSP70 (W27, sc-24, Santa Cruz), HSP90 α/β (F-8, sc-13119, Santa Cruz), IMPDH2 (EPR8365B, ab129165, Abcam), and MTAP (EPR6893, ab126770, Abcam). Membranes were then incubated with peroxidase conjugated secondary antibodies (sc-2004 and sc-2005, Santa Cruz). We used ECL Western Blotting Substrate (Pierce Biotechnology) according to manufacturer’s instructions and the blots were visualized by autoradiography. Quantitative densitometry analysis of western blot bands was performed employing Image J version 10.2 (NIH).

Calvo-Vidal M.N., Zamponi N., Krumsiek J., Stockslager M.A., Revuelta M.V., Phillip J.M., Marullo R., Tikhonova E., Kotlov N., Patel J., Yang S.N., Yang L., Taldone T., Thieblemont C., Leonard J.P., Martin P., Inghirami G., Chiosis G., Manalis S.R, & Cerchietti L. (2021). Oncogenic HSP90 Facilitates Metabolic Alterations in Aggressive B-cell Lymphomas. Cancer Research, 81(20), 5202-5216.

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Characterization of Breast Cancer Cell Lines

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All cell lines were purchased from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). These cell lines were identified by DNA-Fingerprinting, isozyme detection and mycoplasma detection. BT20, MCF-7, MDA-MB-231, MDA-MB-453, MDA-MB-468, HEK 293T, SW527, T47D were cultured in DMEM medium (Gibco, 11965092) plus 10% fetal bovine serum (FBS, Gibco, 10091155) and 1% penicillin/streptomycin (PS, Gibco, 15140163). BT474, BT549 were cultured in RPMI-1640 medium (Gibco, 12633012) plus 10% FBS and 1% PS. DFMO and Putrescine were purchased from Sigma-Aldrich. Antibodies used for western blotting were: Anti-β-actin (1:1000, 4970), Vimentin (1:1000, 5741) and Claudin-1 (1:1000, 13255) from CST, MTAP (1:1000, EPR6893) from Abcam. Antibodies used for IHC analysis were MTAP (1:100, EPR6893) and CD31 (1:100, GB11063-2) from Servicebio.

Zhang Y., Zhang T.T., Gao L., Tan Y.N., Li Y.T., Tan X.Y., Huang T.X., Li H.H., Bai F., Zou C., Pei X.H., Tan B.B, & Fu L. (2022). Downregulation of MTAP promotes Tumor Growth and Metastasis by regulating ODC Activity in Breast Cancer. International Journal of Biological Sciences, 18(7), 3034-3047.

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Western Blot Protein Quantification

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Cells were lysed in protease inhibitor-supplemented RIPA buffer by rotation at 4°C for 30 minutes, followed by centrifugation at 14,000 × g for 5 minutes at 4°C. Protein concentration was determined by BCA assay (Pierce Biotechnology) according to manufacturer's instructions. Protein lysates (10–20 μg) were resolved by SDS-PAGE, transferred to polyvinylidene difluoride (PVDF) membranes, and probed with the indicated primary antibodies: α-tubulin (ab4074, Abcam), β-actin (AC-15, A3854, Sigma), BLNK (2B11, sc-8003, Santa Cruz Biotechnology), CAD (RB18384, AP11110c-ev, Abgent), CLPP (B-12, sc-271284, Santa Cruz Biotechnology), CTPS1 (C-13, sc-131474, Santa Cruz Biotechnology), G6PD (ab76598, Abcam), GAPDH (6C5, MAB374, Millipore Sigma), hexokinase 2 (C-14, sc-6521, Santa Cruz Biotechnology), HSP70 (W27, sc-24, Santa Cruz Biotechnology), HSP90 α/β (F-8, sc-13119, Santa Cruz Biotechnology), IMPDH2 (EPR8365B, ab129165, Abcam), and MTAP (EPR6893, ab126770, Abcam). Membranes were then incubated with peroxidase conjugated secondary antibodies (sc-2004 and sc-2005, Santa Cruz Biotechnology). We used ECL Western Blotting Substrate (Pierce Biotechnology) according to manufacturer's instructions and the blots were visualized by autoradiography. Quantitative densitometry analysis of Western blot bands was performed employing Image J version 10.2 (NIH).

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