NF-kB p65 (RELA) (NM_021975) Human Recombinant Protein

CAT#: TP320780

Recombinant protein of human v-rel reticuloendotheliosis viral oncogene homolog A (avian) (RELA), transcript variant 1

Product images

Western blot analysis of the recombinant NFkBp65 preparation (OriGene TP320780) to check the specificity of two anti-NFkBp65 antibodies: A and C20X. NFkBp65 (25, 50, and 100 ng) was resolved on a NuPAGE 4 - 12% PAGE in MOPs buffer and subjected to Western blot. Figure cited from PLoS ONE, PMID: 25853889

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  Coomassie blue staining of purified RELA protein (Cat# TP320780). The protein was produced from HEK293T cells transfected with RELA cDNA clone (Cat# RC220780) using MegaTran 2.0 (Cat# TT210002).

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  EMSA examines NFkBp65 DNA binding activity. HIV-1 LTR DNA probe was incubated with 100 ng of purified NFkBp65 (OriGene TP320780), and the NFkBp65-DNA complexes were resolved on a 6% non-denaturing polyacrylamide/bisacrylamide gel. NFkBp65-DNA complexes were super-shifted by the addition of anti-NFkBp65 antibody. Anti-NFkBp50 antibody and anti-Myc IgG served as controls. The black arrows indicate the position of the NFkBp65-DNA complex; the grey arrow indicates free HIV-1 LTR DNA probe. The white arrows show the position of the complexes in the presence of the antibody. Figure cited from PLoS ONE, PMID: 25853889

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Specifications

Product Data

• Species: Human
• Expression Host: HEK293T
• Expression cDNA Clone or AA Sequence:
  Protein Sequence

  >RC220780 representing NM_021975
  Red=Cloning site Green=Tags(s)
  
  MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSIPGERSTDTTKTHPTIKINGYTGPG
  TVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELCPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNP
  FQVPIEEQRGDYDLNAVRLCFQVTVRDPSGRPLRLPPVLSHPIFDNRAPNTAELKICRVNRNSGSCLGGD
  EIFLLCDKVQKEDIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPSDREL
  SEPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYP
  FTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQA
  VAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTE
  PMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Tag: C-Myc/DDK
• Predicted MW: 60 kDa
• Concentration: >50 ug/mL as determined by microplate BCA method
• Purity: > 80% as determined by SDS-PAGE and Coomassie blue staining
• Buffer: 25 mM Tris.HCl, pH 7.3, 100 mM glycine, 10% glycerol
• Bioactivity: RELA Activity Verified in a DNA-binding Assay: RELA activity was measured in a colorimetric DNA-binding assay. Double-stranded oligonucleotide containing the RELA consensus DNA-binding sequence was incubated with dilutions of the purified RELA protein. RELA bound to the oligo was captured onto the surface of a microtiter plate and after washing, bound RELA was detected with an anti-RELA primary antibody followed by an HRP-labeled secondary antibody. After initial color development, the reaction was quenched and the color intensity was measured at 450nm.
  ELISA binding assay (PMID: 25584020)
  WB positive control (PMID: 25853889)
  EMSA assay (PMID: 25853889)
  Binding assay (PMID: 26561547)
  Pull-down assay (PMID: 26984196)
• Preparation: Recombinant protein was captured through anti-DDK affinity column followed by conventional chromatography steps.
• Storage: Store at -80°C.
• Stability: Stable for 12 months from the date of receipt of the product under proper storage and handling conditions. Avoid repeated freeze-thaw cycles.

Reference Data

• RefSeq: NP_068810
• Locus ID: 5970
• UniProt ID: Q04206
• Cytogenetics: 11q13.1
• Refseq Size: 1760
• Refseq ORF: 1653
• Synonyms: CMCU; NFKB3; p65
• Summary: NF-kappa-B is a ubiquitous transcription factor involved in several biological processes. It is held in the cytoplasm in an inactive state by specific inhibitors. Upon degradation of the inhibitor, NF-kappa-B moves to the nucleus and activates transcription of specific genes. NF-kappa-B is composed of NFKB1 or NFKB2 bound to either REL, RELA, or RELB. The most abundant form of NF-kappa-B is NFKB1 complexed with the product of this gene, RELA. Four transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Sep 2011]
• Protein Families: Druggable Genome, Transcription Factors
• Protein Pathways: Acute myeloid leukemia, Adipocytokine signaling pathway, Apoptosis, B cell receptor signaling pathway, Chemokine signaling pathway, Chronic myeloid leukemia, Cytosolic DNA-sensing pathway, Epithelial cell signaling in Helicobacter pylori infection, MAPK signaling pathway, Neurotrophin signaling pathway, NOD-like receptor signaling pathway, Pancreatic cancer, Pathways in cancer, Prostate cancer, RIG-I-like receptor signaling pathway, Small cell lung cancer, T cell receptor signaling pathway, Toll-like receptor signaling pathway