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Phospho-NFkB p65 (Ser529) Mouse anti-Human, eFluor® 660, Clone: B33B4WP, eBioscience™ Non-distribution product as customer accommodation.

Mouse Monoclonal Antibody

Manufacturer:  Invitrogen 50986341

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Catalog No. 50-112-4567


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Description

Description

Description: The B33B4WP monoclonal antibody recognizes human NF kappa B (NFkB) p65 subunit when serine 529 is phosphorylated. NFkB, also known as nuclear factor kappa-light chain enhancer of activated B cells, is a ubiquitous transcription factor that regulates the transcription of many genes involved in cell proliferation, apoptosis, development, immunity and cancer. Functional NFkB is a homo- or hetero-dimer composed of 5 members of the NFkB family: p65 (RelA), c-Rel, RelB, p50 (NFkB1, p105 precursor protein), and p52 (NFkB2, p100 precursor protein). The activity of the complex is negatively regulated by binding to IkB inhibitors that sequester NFkB into the cytoplasm, inhibiting its transcriptional activity. NFkB-activating agents like tumor necrosis factor (TNF) alpha, interleukin-1 beta, lipopolysaccharide, camptothecin, and phorbol ester (PMA) induce the phosphorylation and degradation of IkB, leading to the translocation of NFkB to the nucleus where it binds to kB motifs and regulates gene expression. The activity of p65-containing NFkB complexes is positively regulated by phosphorylation of the p65 subunit at serine 529 and/or serine 536. The B33B4WP monoclonal antibody recognizes a single band at 65 kDa in reduced SDS lysates from TNF alpha-stimulated HeLa cells. Applications Reported: This B33B4WP antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This B33B4WP antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of stimulated human peripheral blood cells. This can be used at 5 µL (0.06 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Protocols: We recommend Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins. Alternatively, Protocol C: Two-step protocol: Fixation/Methanol can also be used. Protocol B: One-step protocol: intracellular (nuclear) proteins cannot be used. All Protocols can be found in the "Staining intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online. eFluor™ 660 is a replacement for Alexa Fluor™ 647. eFluor™ 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered. Nuclear factor kB p65 (NF-kB p65) is encoded by the RELA gene and is present on chromosome 11 in humans. NF-kB P65 is also known as RelA (v-rel avian reticuloendotheliosis viral oncogene homolog A) and belongs to the Rel family of proteins. It is one of the two subunits of NF-kB (Nuclear factor kappa-light-chain-enhancer of activated B cells) that heterodimerizes with the other subunits p50 or p52. However, binding of TNF alpha to its cognate receptor phosphorylates IKK which in turn phosphorylates I kB allowing proteasomal degradation of I kB. Phosphorylation of p65 at Ser 536 located in the carboxy-terminal transactivation domains (TAD) leads to nuclear localization of the transcriptionally active complex and NF-kB mediated transactivation of several downstream genes. It specifically plays a key role in transcription of immunoglobulin k (kappa) gene in mature B-lymphoid cells.
Specifications

Specifications

Phospho-NFkB p65 (Ser529)
Monoclonal
5 μL/Test
PBS with 0.1% gelatin, 0.2% BSA and 0.09% sodium azide; pH 7.2
Q04206
nf?b
Mouse
Affinity chromatography
RUO
Antibody
Monoclonal
Human
Flow Cytometry
B33B4WP
eFluor™ 660
RELA
Liquid
RELA
IgG2a, kappa
25 Tests
4° C, store in dark, DO NOT FREEZE!
Primary
5970
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