- NCBI Gene Id:
- Protein Name:
- Transcription factor p65
- Alias Symbols:
- p65, NFKB3, MGC131774, RELA
- Replacement Item:
- This antibody may replace item sc-109 from Santa Cruz Biotechnology.
- Description of Target:
- RABBIT ANTI HUMAN NFkB p65 (pSer536)
- Protein Size (# AA):
- ELISA, IHC-FFPE, WB
- Tissue Tool:
- Find tissues and cell lines supported by DNA array analysis to express RELA.
- RNA Seq:
- Find tissues and cell lines supported by RNA-seq analysis to express RELA.
- The immunogen for anti-RELA antibody: synthetic peptide corresponding to residues surrounding serine 536 of human NFkB p65 protein.
- Predicted Species Reactivity:
- Predicted Homology Based on Immunogen Sequence:
- Product Format:
- Purified IgG - liquid
- Reconstitution and Storage:
- Store at +4oC or at -20oC if preferred.
This product should be stored undiluted.
Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
- Printable datasheet for anti-RELA antibody - OASA08037
- NFkB p65
- Polyclonal IgG
- Application Info:
- Immunohistology - Paraffin: 1/50 - 1/250
ELISA: 1/500 - 1/3000
Western Blotting(1): 1/100 - 1/1000
(1) Special conditions apply. See *
- Additional Information:
- Histology Positive Control Tissue: Human kidney distal tubules and collecting ducts.
- Preservative Stabilisers: 0.09% Sodium Azide (NaN3)
Antiserum Preparation: Antisera to NFkB p65 (pSer536) were raised by repeated immunisation of rabbits with highly purified antigen. Purified IgG was prepared from whole serum by affinity chromatography.
- Approx Protein Conc: IgG concentration 0.5mg/ml
Buffer Solutions: Phosphate buffered saline pH7.2
- Protocol Information:
- Citation: 1: Yamamoto K, Arakawa T, Ueda N, Yamamoto S. Transcriptional roles of nuclearfactor kappa B and nuclear factor-interleukin-6 in the tumor necrosis factoralpha-dependent induction of cyclooxygenase-2 in MC3T3-E1 cells. J Biol Chem.1995 Dec 29;270(52):31315-20. PubMed PMID: 8537402.
Experiment Name: Electrophoretic mobility shift assay targeting the NFkB site
Experiment Background: When a mouse osteoblastic cell line MC3T3-E1 was cultured in the presence of tumor necrosis factor a (TNFa), the release of prostaglandin E2 and the cyclooxygenase activity increased in a dose- and time-dependent manner. The increase of the enzyme activity was attributed mostly to the induction of cyclooxygenase-2 rather than cyclooxygenase-1 as judged by the inhibitory effect of NS398, Western blotting, and Northern blotting. In this system Kei et al. attempted to elucidate the transcriptional regulation of the cyclooxygenase-2 gene. As examined by the luciferase assay, two positive regulatory regions (2186 to 2131 and 2512 to 2385 base pairs) were found in the 5’-flanking promoter region of the mouse cyclooxygenase-2 gene in the TNFa-stimulated cells. The former included putative NF-IL6 (C/EBPb) and AP2 elements, and the latter contained the NFkB motif. A DNA probe including the NF-IL6 and AP2 sites gave positive bands upon electrophoretic mobility shift assay using the nuclear extracts of MC3T3-E1 cells. The bands were supershifted by the addition of anti-NFIL6 antibody but not by anti-AP2 antibody. A probe including the NFkB site also gave positive bands, which were supershifted by anti-NFkB p50 and p65 antibodies. Furthermore, when the motif of NF-IL6 or NFkB or both was subjected to point mutation, the luciferase activity was markedly reduced. These data suggested a potential role of both NF-IL6 and NFkB in the induction of cyclooxygenase-2 by TNFa.
Experimental Steps: Probe g1 was incubated for 0.5 h with the nuclear extracts (2.3 mg protein) of MC3T3-E1 cells treated with 0 ng/ml (lane 3), 0.2 ng/ml (lane 4), 2 ng/ml (lane 5), and 20 ng/ml (lanes 6 and 9) TNFa. The nuclear extracts before the addition of TNFa were also incubated with g1 (lane 2). Anti-NFkB p50 (lane 10) or anti-NFkB p65 (lane 11) antibody was incubated together with the nuclear extracts from the cells stimulated with 20 ng/ml TNFa. Purified NFkB p50 protein was incubated with probe g1 (lane 7). Probe g2 with a mutation for NFkB was incubated with the nuclear extracts from the cells stimulated with 20 ng/ml TNFa (lanes 13 and 14). Anti-NFkB p50 antibody was also present (lane 14). Lanes 1, 8, and 12 were control runs without the nuclear extracts.
Other Reagents Used: Phenylmethylsulfonyl fluoride, dithiothreitol. KCl, MgCl2, EDTA
Number Of Protocols: 1
- Target Reference:
- 1. Mattioli, I. et al. (2004) Transient and selective NF-kappa B p65 serine 536 phosphorylation induced by T cell costimulation is mediated by I kappa B kinase beta and controls the kinetics of p65 nuclear import. J. Immunol. 172: 6336-6344.