RelA/NFkB p65 Antibody (PSH0-27)

Western Blot: RelA/NFkB p65 Antibody (PSH0-27) [NBP3-32648] - Western blot analysis of RelA/NFkB p65 on different lysates with Rabbit anti-RelA/NFkB p65 antibody (NBP3-32648) at 1/500 dilution. Lane 1: Hela cell lysate (control) Lane 2: Hela-RELA-KO (1) Lane 3: Hela-RELA-KO (2) Lysates/proteins at 35 ug/Lane. Predicted band size: 60 kDa Observed band size: 65 kDa Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32648) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: RelA/NFkB p65 Antibody (PSH0-27) [NBP3-32648] - Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-RelA/NFkB p65 antibody (NBP3-32648) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32648) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry/ Immunofluorescence: RelA/NFkB p65 Antibody (PSH0-27) [NBP3-32648] - Immunocytochemistry analysis of NIH/3T3 cells treated with or without 50μg/mL TNF-alpha for 20 minutes labeling RelA/NFkB p65 with Rabbit anti-RelA/NFkB p65 antibody (NBP3-32648) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-RelA/NFkB p65 antibody (NBP3-32648) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Flow Cytometry: RelA/NFkB p65 Antibody (PSH0-27) [NBP3-32648] - Flow cytometric analysis of NIH/3T3 cells labeling RelA/NFkB p65. Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32648, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

• Reactivity: Hu, Mu, Rt, Pm
• Applications: WB, Flow, ICC/IF, IHC
• Clone: PSH0-27
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

RelA/NFkB p65 Antibody (PSH0-27) Summary

• Description: Novus Biologicals Rabbit RelA/NFkB p65 Antibody (PSH0-27) (NBP3-32648) is a recombinant monoclonal antibody validated for use in IHC, WB, Flow and ICC/IF. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Synthetic peptide within human RELA aa 502-551 / 551. (Uniprot: Q04206)
• Localization: Nucleus, Cytoplasm.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: RELA
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1ug/mL
  • Immunocytochemistry/ Immunofluorescence 1:200
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:1000
  • Western Blot 1:1000
• Theoretical MW: 65 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH7.4), 0.1% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for RelA/NFkB p65 Antibody (PSH0-27)

• MGC131774
• nf kb p65
• NFkB p65
• NF-kB p65
• NFKB3
• NFKB3v-rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor ofkappa light polypeptide gene enhancer in B-cells 3 (p65))
• Nuclear factor NF-kappa-B p65 subunit
• Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3transcription factor p65
• p65
• p65RelA
• rela p65
• RelA
• v-rel reticuloendotheliosis viral oncogene homolog A (avian)
• v-rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappalight polypeptide gene enhancer in B-cells 3, p65

Background

Nuclear factor B (NF-B) encompasses an important family of inducible transcriptional activators that regulate a wide variety of cellular and viral genes. The family members include p50, p52, p65 (RelA), c-Rel, and RelB. The Nf-kB p65 subunit, similar to two others in the B family, RelB and c-Rel, contains two transactivation domains in the C-terminal region of the protein. Association with inhibitory proteins of the I B family, retains NF- B in the cytoplasm. Degradation of IB proteins exposes the nuclear localization sequence (NLS), leading to nuclear translocation and subsequent binding of NF-B to DNA. In addition to the nuclear translocation and DNA binding, Nf-kB's transcriptional activity is also regulated by coactivators and corepressors in the nucleus. Interaction of p65 with coactivators; CREB-binding protein and p300, increases its transcriptional activity.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: RELA