NFκB p65 antibody (mAb)

Application Notes

Applications Validated by Active Motif:
WB: 2 - 5 µg/ml dilution

For optimal results, primary antibody incubations should be performed at room temperature. The addition of 0.1% Tween 20 to all blocking solutions may also reduce background. Individual optimization may be required.

Published Applications

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

ChIP

View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.

Immunogen

This NFkB p65 antibody was raised against a synthetic peptide corresponding to amino acid residues 526-539 of human NFκB p65.

Buffer

PBS containing 0.05% BSA and 0.05% sodium azide. Sodium azide is highly toxic.

Background

NFκB p65 (Nuclear factor NF-kappa-B p65 subunit also known as NFκB p65 or RelA) is a member of the Rel family of transcription factors that are involved in processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. The Rel family includes c-Rel, RelA (NFκB p65, NFκB3) and RelB (I-Rel). Rel proteins form dimers with the NFκB p50 (proteolytically processed from p105/NFκB1) and p52 (proteolytically processed from p100/NFκB2) members of the NFκB family that bind DNA and activate transcription. The NFκB p65/p50 heterodimer is the most abundant form of NFκB.

NFκB signaling is negatively regulated by the sequestration of the NFκB complex in the cytoplasm by its association with the IκB family of inhibitory proteins. The IκB Kinase (IKK) complex is the key enzyme involved in the activation and translocation of NFκB. The IKK complex is composed of two catalytic subunits (IKKα and IKKβ) and a regulatory subunit (IKKγ). Upon its activation, the IKK complex phosphorylates IκB proteins (IκBα, IκBα phospho Ser32, 36), which marks them for ubiquitination and degradation. This enables the NFκB complex to translocate to the nucleus where it regulates transcription by binding to DNA.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT^® High Sensitivity Kit or our magnetic bead-based ChIP-IT^® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with NFκB p65 antibody (mAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Store at 4°C for short term. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

• ChIP = Chromatin Immunoprecipitation;
• ChIP = ChIP Sequencing;
• CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
• CUT&Tag = Cleavage Under Targets and Tagmentation;
• DB = Dot Blot;
• ELISA = Enzyme-linked Immunosorbent Assay;
• EMSA = Electrophoretic Mobility Shift Assay
• FC = Flow Cytometry;
• ICC = Immunocytochemistry;
• IF = Immunofluorescence;
• IHC = Immunohistochemistry;
• IP = Immunoprecipitation;
• MeDIP = Methyl-DNA Immunoprecipitation;
• NEU = Neutralizing;
• WB = Western Blotting