Rabbit anti nf κb p65 d14e12

Manufactured by Cell Signaling Technology

Rabbit anti-NF-κB p65 (D14E12) is a primary antibody that recognizes the p65 subunit of the NF-κB transcription factor. It is intended for use in various research applications such as Western blotting, immunoprecipitation, and immunohistochemistry.

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NF-κB p65 (724 citations) NF-κB (393 citations) Anti-NF-κB p65 (293 citations) Anti-NF-κB (140 citations) Rabbit anti-NF-κB p65 (60 citations) Rabbit anti-p65 (44 citations) Rabbit anti-NF-κB (16 citations) Anti-NF-κB p65 (D14E12, (10 citations) NF-κB p65 (D14E12) (10 citations) Anti-p65 (D14E12, (7 citations)

4 protocols using rabbit anti nf κb p65 d14e12

Molecular Mechanisms of Endoplasmic Reticulum Stress

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Primary antibodies used were as follows: mouse anti-XBP1s (Biolegend, 647502), rabbit anti-IRE1 (Cell Signaling, 3294), rabbit anti-PERK (Cell Signaling Technology, 3192), rabbit anti-eIF2α (Cell Signaling Technology, 5324), rabbit anti-p(S51)-eIF2α (Cell Signaling Technology, 3398), mouse anti-ATF6 (CosmoBio, BAM-73-500-EX), rabbit anti-NLRP3 D2P5E (Cell signaling technology, 13158), rabbit anti-NF-κB p65 (D14E12) (Cell signaling technology, 8242), mouse anti-IL-1β (R&D Systems, MAB601), rabbit anti-ASC (Santa Cruz, sc-22514-R), rabbit anti-caspase 1 (Santa Cruz, sc-622), rabbit anti-caspase-1 p10 (Santa Cruz, sc-515), TXNIP (Santa Cruz, sc-166234), and rabbit anti-Actin (Sigma, A2066). Secondary antibodies were horseradish peroxidase-tagged goat anti-mouse (Jackson Laboratories, 115-035-003) and goat anti-rabbit antibodies (Jackson Laboratories, 111-035-003). Tunicamycin (T7765), Phorbol 12-myristate 13-acetate (PMA) (P8139), LPS (L2630), and ATP (A6419) were purchased from Sigma-Aldrich while nigericin (tlrl-nig) was obtained from Invivogen. IRE1 inhibitor MKC8866 was provided by Fosun Orinove.

Talty A., Deegan S., Ljujic M., Mnich K., Naicker S.D., Quandt D., Zeng Q., Patterson J.B., Gorman A.M., Griffin M.D., Samali A, & Logue S.E. (2019). Inhibition of IRE1α RNase activity reduces NLRP3 inflammasome assembly and processing of pro-IL1β. Cell Death & Disease, 10(9), 622.

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Quantitative IHC Analysis of DNA Damage and Inflammation Markers

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IHC analyses were conducted using 5-µm paraffin sections as described before^{23 (link),41 (link)}. Antibodies used are as follows: mouse anti-γH2A.X (Ser139) (clone JBW301, Millipore Sigma, 05-636, 1:200), mouse anti-8-oxo-dG (Trevigen, 4354-MC-050, 1:500), rabbit anti-NFκB p65 (D14E12, Cell Signaling, 8242S, 1:1000), and rabbit anti-phospho-NFκB p65 (Ser536) (Abcam, AB86299, 1:500). Immunoreactivity was evaluated, and the number of positive ductal epithelial cells was scored and converted to the percentage of all ductal epithelial cells. Three mice per genotype (one abdominal mammary gland per mouse and 5–10 ducts per mammary gland) were analyzed for each genotype and each marker.

Huo Y., Selenica P., Mahdi A.H., Pareja F., Kyker-Snowman K., Chen Y., Kumar R., Da Cruz Paula A., Basili T., Brown D.N., Pei X., Riaz N., Tan Y., Huang Y.X., Li T., Barnard N.J., Reis-Filho J.S., Weigelt B, & Xia B. (2021). Genetic interactions among Brca1, Brca2, Palb2, and Trp53 in mammary tumor development. NPJ Breast Cancer, 7, 45.

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Western Blotting Antibody Panel for Cellular Signaling

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The following antibodies were used for western blotting: rabbit anti-phospho-p38 MAPK (Thr180/Tyr182) (D3F9) (#9211; Cell Signaling), rabbit anti-p38 MAPK (#9212; Cell Signaling), rabbit anti-phospho-NF-κB p65 (Ser536) (93H1) (#3033; Cell Signaling), rabbit anti-NF-κB p65 (D14E12) (#8242; Cell Signaling), rabbit anti-phospho-MAPKAPK-2 (Thr334) (27B7) (#3007; Cell Signaling), rabbit anti-MAPKAPK-2 (#3042; Cell Signaling), rabbit anti-active caspase-1 (#4199; Cell Signaling), rabbit anti-caspase-1 (#3866; Cell Signaling), mouse anti-caspase-8 (8CSP03) (sc-56070; Santa Cruz Biotechnology), rabbit anti-caspase-5 (D3G4W) (#46680; Cell Signaling), and mouse anti-β-actin (8H10D10) (#3700; Cell Signaling). All primary antibodies were diluted 1:1000. Secondary antibodies used were goat anti-rabbit IgG, HRP-linked antibody (#7074; Cell Signaling) and horse anti-mouse IgG, HRP-linked antibody (#7076; Cell Signaling), and they were diluted to 1:5000. Blots were developed with an enhanced chemiluminescence substrate (Thermo Scientific) and bands were identified by exposure of the membrane to X-ray film (Kodak, CareStream Health).

Phuong M.S., Hernandez R.E., Wolter D.J., Hoffman L.R, & Sad S. (2021). Impairment in inflammasome signaling by the chronic Pseudomonas aeruginosa isolates from cystic fibrosis patients results in an increase in inflammatory response. Cell Death & Disease, 12(3), 241.

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Western Blot Analysis of Signaling Pathways

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Tumor cells with or without TNF-α treatment or genetic modification were lysed in RIPA lysis buffer with protease and phosphatase inhibitor cocktail (Thermo Scientific, 78444). Equivalent amounts of protein from whole cell lysates were separated by SDS-PAGE and transferred onto PVDF membranes (Bio-Rad). Membranes were blocked in 5% nonfat milk in PBS plus 0.1 % Tween-20 and stained with primary antibodies, followed by probing with horseradish peroxidase-conjugated secondary antibodies (Jackson Immunoresearch). Primary antibodies used include goat anti-IRF6 (Novus Biologicals, NBP1–51911), rabbit anti-TRADD (Cell Signaling Technology, 3694), mouse anti-FADD (1F7; Enzo Life Sciences, ADI-AAM-212-E), rabbit anti-Caspase-8 (D35G2; Cell Signaling Technology, 4790), rabbit anti-kBα (Cell Signaling Technology, 9242), rabbit anti-phospho-NF-ΚB p65 (Ser536) (Cell Signaling Technology, 3031), rabbit anti-NF-ΚB p65 (D14E12; Cell Signaling Technology, 8242), and rabbit anti-GAPDH (14C10; Cell Signaling Technology, 2118). ECL solution (Thermo Scientific, 32106) was used as a substrate and band signals were detected using ChemiDoc (Bio-Rad).

Kim I.K., Diamond M., Yuan S., Kemp S., Li Q., Lin J., Li J., Norgard R., Thomas S., Merolle M., Katsuda T., Tobias J., Politi K., Vonderheide R, & Stanger B. (2023). Plasticity-induced repression of Irf6 underlies acquired resistance to cancer immunotherapy. Research Square.

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