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RelA/NFkB p65 Antibody (PSH0-27)

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-32648

Recombinant Monoclonal Antibody
Novus Biologicals, part of Bio-Techne
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NBP3-32648

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Monkey

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # PSH0-27

Concentration

1 mg/ml

Product Summary for RelA/NFkB p65 Antibody (PSH0-27)

Immunogen

Synthetic peptide within human RELA aa 502-551 / 551. (Uniprot: Q04206)

Localization

Nucleus, Cytoplasm.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

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.

Scientific Data Images for RelA/NFkB p65 Antibody (PSH0-27)

RelA/NFkB p65 Antibody (PSH0-27)

Western Blot: RelA/NFkB p65 Antibody (PSH0-27) [NBP3-32648]

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.
RelA/NFkB p65 Antibody (PSH0-27)

Immunohistochemistry: RelA/NFkB p65 Antibody (PSH0-27) [NBP3-32648]

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.
RelA/NFkB p65 Antibody (PSH0-27)

Immunocytochemistry/ Immunofluorescence: RelA/NFkB p65 Antibody (PSH0-27) [NBP3-32648]

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.

Applications for RelA/NFkB p65 Antibody (PSH0-27)

Application
Recommended Usage

Flow Cytometry

1ug/mL

Immunocytochemistry/ Immunofluorescence

1:200

Immunohistochemistry-Paraffin

1:1000

Western Blot

1:1000
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH7.4), 0.1% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: RelA/NFkB p65

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.

Long Name

v-rel Reticuloendotheliosis Viral Oncogene Homolog A

Alternate Names

NFkB p65, NFKB3, p65RelA

Gene Symbol

RELA

Additional RelA/NFkB p65 Products

Product Documents for RelA/NFkB p65 Antibody (PSH0-27)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for RelA/NFkB p65 Antibody (PSH0-27)

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.

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