Anti-NF-kB p65/RELA Antibody

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Datasheet

Protocols

COA

MSDS

SKU	PA1125
Size	100μg/vial
Reactivity	Human, Mouse, Rat
Clonality	Polyclonal
Host	Rabbit
Ig Isotype	N/A
Applications	WB

• Overview
• Assay Details
• Target Info
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Overview

Product Name	Anti-NF-kB p65/RELA Antibody
SKU/Catalog Number	PA1125
Storage & Handling	At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.
Size	100μg/vial
Description	Rabbit IgG polyclonal antibody for Transcription factor p65(RELA) detection. Tested with WB in Human;Mouse;Rat.
Cite This Product	Anti-NF-kB p65/RELA Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1125)
Host	Rabbit
Contents/Buffer	Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
Form	Lyophilized
Immunogen	A synthetic peptide corresponding to a sequence at the N-terminus of human NF-kB p65(116-131aa AISQRIQTNNNPFQVP), identical to the related rat sequence, and different from the related mouse sequence by one amino acid.
Reactivity	Human, Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat

Boster's Secondary Antibodies And IHC, WB Kits

The following reagents are used to generate the images below.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.

Images And Assay Conditions

Anti-NF-kB p65 antibody, PA1125, Western blotting
All lanes: Anti NF-kB p65 (PA1125) at 0.5ug/ml
Lane 1: Human Colon Cancer Tissue Lysate at 50ug
Lane 2: HELA Whole Cell Lysate at 40ug
Predicted bind size: 65KD
Observed bind size: 65KD

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id	Q04206
Gene Name	RELA
Protein Name	Transcription factor p65
Alternative Names	Transcription factor p65;Nuclear factor NF-kappa-B p65 subunit;Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3;RELA;NFKB3;
Subcellular Localization	Nucleus. Cytoplasm. Colocalized with DDX1 in the nucleus upon TNF-alpha induction (By similarity). Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalizes with GFI1 in the nucleus after LPS stimulation. .
Molecular Weight	60219 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function	NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells (PubMed:15790681). .
Research Areas	Human, Mouse, Rat *You can search these to find other products in these research areas.
Background	The p65(RELA) heterodimer is the most abundant form of NFKB. This gene is located on 11q13, which consists of 10 exons and spans about 8.1 kb of DNA. In rat sciatic nerves, the expression of the activated p65 subunit of NFKB was high in the nuclei of premyelinating Schwann cells and then progressively declined until it was nearly absent in adults. The transcriptional activity of NF-kappa-B is stimulated upon phosphorylation of its p65 subunit on serine-276 by protein kinase A(PKA). The transcriptional coactivator CBP/p300 associates with NF-kappa-B p65 through 2 sites, an N-terminal domain that interacts with the C-terminal region of unphosphorylated p65, and a second domain that only interacts with p65 phosphorylated on serine-276.

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Polyclonal antibody for NFkB p65/RELA detection. Host: Rabbit.Size: 100μg/vial. Tested applications: WB. Reactive species: Human. NFkB p65/RELA information: Molecular Weight: 60219 MW; Subcellular Localization: Nucleus. Cytoplasm. Colocalized with DDX1 in the nucleus upon TNF-alpha induction (By similarity). Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalizes with GFI1 in the nucleus after LPS stimulation.

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PA1125

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Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)

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Troubleshooting

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Customer Q&As

• Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
  A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
• Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WB
  A: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
• Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
  A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
• Q: What should I use for negative control?
  A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
• Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
  A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
• Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
  A: You can find the immunogen sequence under "
• Q: What is the expected band size? Why is it different than the observed band size?
  A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
• Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
  A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
• Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
  A: Check our protocols under the tech support tab.
• Q: What are some alternative names that could be used to describe this product?
  A: Some common names include but are not limited to p65 antibody, rela antibody