Anti-NF-kB p65/RELA Antibody

Product Name

Anti-NF-kB p65/RELA Antibody

View all RELA Antibodies

SKU/Catalog Number

PA1669

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-NF-kB p65/RELA Antibody catalog # PA1669. Tested in IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-NF-kB p65/RELA Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1669)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence in the middle region of human NF-kB p65, identical to the related rat and mouse sequences.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PA1669 is reactive to RELA in Human, Mouse, Rat

Applications

PA1669 is guaranteed for IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

65 kDa

Calculated molecular weight

60.219kDa

Background of RELA

RELA (V-REL AVIAN RETICULOENDOTHELIOSIS VIRAL ONCOGENE HOMOLOG A), also called NFKB3 or NFKB, p65 SUBUNIT. NFKB1 or NFKB2 is bound to REL, RELA, or RELB to form the NFKB complex. The NFKB complex is inhibited by I-kappa-B proteins, which inactivate NFKB by trapping it in the cytoplasm. The p65 (RELA) heterodimer is the most abundant form of NFKB. And the RELA gene is located on 11q13.1. RELA is a nonhistone substrate of HDAC3 and that IKBA-dependent nuclear export of the HDAC3-deacetylated RELA replenishes the depleted cytoplasmic pool of latent NFKB-IKBA complexes for subsequent NFKB responses. RELA nucleocytoplasmic redistribution coincided with export of PPARG, and immunoprecipitation analysis indicated that PPARG-RELA association was dependent on the PPARG C-terminal ligand-binding domain. IKK-dependent phosphorylation of RELA on ser468 enhanced binding of GCN5 to RELA and RELA ubiquitination.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Immunocytochemistry/Immunofluorescence , 5μg/ml, Human, -

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of NF-kB p65 using anti-NF-kB p65 antibody (PA1669).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human Caco-2 whole cell lysates,
Lane 4: human Raji whole cell lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: rat C6 whole cell lysates,
Lane 7: mouse NIH/3T3 whole cell lysates,
Lane 8: mouse Ana-1 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NF-kB p65 antigen affinity purified polyclonal antibody (Catalog # PA1669) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for NF-kB p65 at approximately 65 kDa. The expected band size for NF-kB p65 is at 60 kDa.

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Figure 2. IHC analysis of NF-kB p65 using anti-NF-kB p65 antibody (PA1669).
NF-kB p65 was detected in a paraffin-embedded section of Human Mammary Cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-NF-kB p65 Antibody (PA1669) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. IF analysis of NF-kB p65 using anti-NF-kB p65 antibody (PA1669).
NF-kB p65 was detected in an immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-NF-kB p65 Antibody (PA1669) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Gene/Protein Information For RELA (Source: Uniprot.org, NCBI)

Alternative Names

NF kB p65; NFkB p65; NF-kB p65; p65; rela p65; RelA; 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 flow cytometry; p65 immunohistochemistry; p65; p65RelA; rela p65; RelA; v-rel reticuloendotheliosis viral oncogene homolog A (avian); v-rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappalight po RELA CMCU, NFKB3, p65 RELA proto-oncogene, NF-kB subunit transcription factor p65|NF-kappa-B p65delta3|NF-kappa-B transcription factor p65|nuclear factor NF-kappa-B p65 subunit|nuclear factor of kappa light polypeptide gene enhancer in B-cells 3|v-rel avian reticuloendotheliosis viral oncogene homolog A

For more info on RELA, check out the RELA Infographic

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In this infographic, you will see the following information for RELA: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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