Phospho-NFkB p65 (Ser529) Monoclonal Antibody (B33B4WP), eFluor 660

货号： 50-9863-41,50-9863-42

规格：25 Tests,100 Tests

价格：1237,3202

产品类型：流式抗体

品牌：Thermo Fisher

物种：人

宿主：小鼠

抗体亚型：IgG2a, kappa

克隆号：B33B4WP

荧光染料：eFluor 660

类型:	单抗	同型对照：	Mouse IgG2a kappa Isotype Control (eBM2a), eFluor 660
浓度：	5 µL/Test	用法：	5 µL (0.06 µg)/test(Flow)

产品详细信息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.靶标信息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.

数据

Phospho-NFkB p65 (Ser529) Antibody (50-9863-42) in FlowIntracellular staining of unstimulated (left) or 15-minute PMA/ionomycin-stimulated (right) normal human peripheral blood cells with Anti-Human CD57 PE (Product # 12-0577-42) and Anti-Human phospho-NF kappa B p65 (S529) eFluor® 660, using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Cells in the lymphocyte gate were used for analysis.

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