Phospho-NFkB p65 (Ser529), eFluor 660, clone: B33B4WP, eBioscience™
Mouse Monoclonal Antibody
Brand: Affymetrix eBioscience 50-9863-41
Code : NEW
Additional Details : Weight : 0.23750kg
DescriptionDescription: 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 65kDa 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: 0.2μm post-manufacturing filtered.
|Phospho-NF kappa B p65 (S529)|
|PBS with 0.1% gelatin, 0.2% BSA and 0.09% sodium azide; pH 7.2|
|Nuclear factor kappa B p65 subunit, RelA|
|Store at 2-8°C. Do not freeze. Light-sensitive material.|
For Research Use Only.