Anti phospho nf kb p65 ser536

Manufactured by Cell Signaling Technology

Sourced in China

Anti-phospho-NF-kB p65 (Ser536) is a primary antibody that recognizes the phosphorylated form of the NF-kB p65 subunit at serine 536. This antibody can be used to detect and quantify the activated form of NF-kB p65 in various experimental systems.

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Lab products found in correlation

Anti nf kb p65, by Cell Signaling Technology (2 mentions) Protein assay kit, by Thermo Fisher Scientific (1 mentions) Anti gapdh, by Novus Biologicals (1 mentions) Anti nrf2, by Cell Signaling Technology (1 mentions) Anti il 6, by Abcam (1 mentions) Anti irf3, by Cell Signaling Technology (1 mentions) Anti sting, by Proteintech (1 mentions) Anti cd63, by ABclonal (1 mentions) Anti alix 12422 1 ap, by Proteintech (1 mentions) Anti cd81 66866 1 ig, by Proteintech (1 mentions) Anti phospho sting ser366, by Cell Signaling Technology (1 mentions) Anti phospho sting ser365, by Cell Signaling Technology (1 mentions) Anti nf kb p65, by ABclonal (1 mentions) Anti phospho irf3 ser396, by Cell Signaling Technology (1 mentions) Anti β actin, by Cell Signaling Technology (1 mentions) Anti gapdh, by Beyotime (1 mentions) Anti calnexin, by Abcam (1 mentions) Protease inhibitor cocktail, by Keygen Biotech (1 mentions) Pierce bca protein assay kit, by Keygen Biotech (1 mentions) Phosphatase inhibitor cocktail, by Keygen Biotech (1 mentions)

3 protocols using anti phospho nf kb p65 ser536

Cellular Protein Analysis via WES

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Cell protein was extracted using RIPA lysis buffer containing protease and phosphatase inhibitor cocktails (cat. no. 539134, Millipore and cat. no. 78420, Thermo Fisher Scientific), and the protein concentration was determined using the protein assay kit (cat. no. 23200, Thermo Fisher Scientific, United States). Separation of proteins was subjected to the WES system (ProteinSimple, United States) and 12–230 kDa separation modules were used. For target proteins detection, the following antibodies were used: anti-NF- kB p65 (Cell Signaling Technology, #6956), anti-phospho- NF- kB p65 (Ser536) (Cell Signaling Technology, #3033), anti-Nrf2 (Cell Signaling Technology, #12721), anti-IL-6 (Abcam, ab9324), and anti-GAPDH (Novus Biologicals, NB300-325).

Chen Y., Wang B., Lai W.F., Chen Y., Pan R., Tang Z, & Liu D. (2022). Chinese herbal formula (GCNY)-medicated serum alleviates peroxidation induced by H2O2 in human microglial cells. Frontiers in Neuroscience, 16, 990040.

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Proteomic Analysis of Colon Tissue Fractions

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Colon tissues, cells, and EVs fractions were homogenized using the RIPA buffer (Jiangsu KeyGEN BioTECH, Nanjing, China), phosphatase inhibitor cocktail, and protease inhibitor cocktail (Jiangsu KeyGEN BioTECH, Nanjing, China). Pierce BCA Protein Assay Kit (Jiangsu KeyGEN BioTECH, Nanjing, China) was used to determine the protein concentration. Anti-CD63 (A5271, ABclonal), anti-Alix (12422-1-AP, Proteintech), anti-CD81 (66866-1-Ig, Proteintech), anti-Calnexin (ab22595, Abcam), anti-STING (19851-1-AP, Proteintech), anti-Phospho-STING (Ser366) (85735, Cell Signaling Technology), anti-Phospho-STING (Ser365) (72971, Cell Signaling Technology), anti-IRF3 (4302, Cell Signaling Technology), anti-Phospho-IRF3 (Ser396) (29047, Cell Signaling Technology), anti-NF-kB p65 (A19653, ABclonal), anti-Phospho-NF-kB p65 (Ser536) (3303, Cell Signaling Technology), anti-Beta-Actin (4970, Cell Signaling Technology), anti-GAPDH (AF1186, Beyotime Biotechnology) and secondary antibodies (KGP1201, Jiangsu KeyGEN BioTECH, Nanjing, China) were used for western blot.

Zhao F., Zheng T., Gong W., Wu J., Xie H., Li W., Zhang R., Liu P., Liu J., Wu X., Zhao Y, & Ren J. (2021). Extracellular vesicles package dsDNA to aggravate Crohn’s disease by activating the STING pathway. Cell Death & Disease, 12(9), 815.

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NF-κB Signaling Pathway Analysis

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Aliquots containing 1 3 10 6 cells were washed with PBS and lysed in RIPA buffer (150 mmol/L NaCl, 1.0% NP-40, 0.1% SDS, 0.1% sodium deoxycholate, 5 mmol/L EDTA, and 10 mmol/L Tris-HCl, pH 7.4) containing protease inhibitors. Proteins were separated by using SDS-PAGE and transferred to membranes, and blots were probed with the following primary antibodies: anti-TNFAIP3, anti-phospho-IKKa/b Ser176/180, anti-IKKa, anti-phospho-IkBa Ser32, anti-IkBa, anti-phospho-NF-kB p65 Ser536, anti-NF-kB p65, anti-phospho-p38 mitogen-activated protein kinase (MAPK), anti-p38 MAPK, anti-SMC1 (all from Cell Signaling, Danvers, Mass), anti-glyceraldehyde-3-phosphate dehydrogenase, anti-CASP3 (GeneTex, Irvine, Calif), anti-green fluorescent protein (GFP; TaKaRa Clontech, Siga, Japan), or anti-b-actin (Sigma-Aldrich, St Louis, Mo). Primary antibodies were detected with horseradish peroxidase-conjugated anti-rabbit or anti-mouse secondary antibody (GE Healthcare, Little Chalfont, United Kingdom).

Takagi M., Ogata S., Ueno H., Yoshida K., Yeh T., Hoshino A., Piao J., Yamashita M., Nanya M., Okano T., Kajiwara M., Kanegane H., Muramatsu H., Okuno Y., Shiraishi Y., Chiba K., Tanaka H., Bando Y., Kato M., Hayashi Y., Miyano S., Imai K., Ogawa S., Kojima S, & Morio T. (2017). Haploinsufficiency of TNFAIP3 (A20) by germline mutation is involved in autoimmune lymphoproliferative syndrome. The Journal of allergy and clinical immunology, 139(6).

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