Phospho-IkB alpha (Ser32, Ser36) Monoclonal Antibody (RILYB3R), PE

货号：12-9035-41,12-9035-42

规格：25 Tests,100 Tests

价格：1299,3217

产品类型：流式抗体

品牌：Thermo Fisher

物种：人/小鼠

宿主：小鼠

抗体亚型：IgG2b, kappa

克隆号：RILYB3R

荧光染料：PE

类型:	单抗	同型对照：	Mouse IgG2b kappa Isotype Control (eBMG2b), PE
浓度：	5 µL/Test	用法：	5 µL (0.125 µg)/test(Flow)

产品详细信息Description: This RILYB3R monoclonal antibody recognizes human and mouse nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor, alpha (I kappa B alpha) when phosphorylated on serines 32 and 36 (S32/S36). I kappa B alpha is one member of a family of cellular proteins that functions to inhibit classical/canonical NF-kappa B signaling by masking the nuclear localization signals (NLS) of NF-kappa B proteins and keeping them sequestered in an inactive state in the cytoplasm. Classical/canonical NF-kappa B signaling is initiated in response to myriad stimuli including engagement of T cell and B cell receptors, growth factors, and inflammatory stimuli (reactive oxygen species, TNF alpha, IL-1) and results in the activation of the I kappa B kinase (IKK) complex that includes IKK alpha, IKK beta, and NEMO. IKK phosphorylates I kappa B alpha resulting in its ubiquitination, degradation, and subsequent translocation of NF-kappa B transcription factor proteins into the nucleus.Applications Reported: This RILYB3R antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This RILYB3R antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.125 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.Staining Protocol: Protocol A and Protocol C are recommended for this monoclonal antibody. Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to "Clone Performance Following Fixation/Permeabilization" located in the Best Protocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online.Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.Filtration: 0.2 µm post-manufacturing filtered.靶标信息IkB-alpha is a 40 kDa protein that functions to inhibit NF- kappaB activity. The inhibition occurs via protein-protein interaction between I kappaB proteins and NF- kappaB dimers in the cytosol. The interaction of I kappa B-alpha with NF- kappaB masks the nuclear localization sequence of NF- kappaB, preventing NF- kappaB translocation to the nucleus. A variety of stimuli can activate gene expression by liberating NF- kappaB through the degradation of I kappaB alpha. These stimuli include the proinflammatory cytokines TNF- alpha and IL-1 beta, chemokines, PMA, growth factors, LPS, UV irradiation, viral infection, as well as various chemical and physical stresses. In humans, the gene is located on the q arm of chromosome 14. Activation of NFkB requires that IkB be phosphorylated on specific serine residues, which results in targeted degradation of IkB. IkB kinase alpha (IKK alpha), previously designated CHUK, interacts with IkB-alpha and specifically phosphorylates IkB-alpha on the sites that trigger its degradation Serines 32 and 36. IKK alpha appears to be critical for NFkB activation in response to proinflammatory cytokines. Phosphorylation of IkB by IKK alpha is stimulated by the NFkB inducing kinase (NIK), which itself is a central regulator for NFkB activation in response to TNF and IL-1. The functional IKK complex contains three subunits, IKK alpha, IKK beta and IKK gamma, and each appear to make essential contributions to IkB phosphorylation.

数据

Phospho-IkB alpha (Ser32, Ser36) Antibody (12-9035-42) in FlowIntracellular staining of untreated (blue histogram) or 60-minute Human TNF alpha Recombinant Protein- (Product # 14-8329-81) and Calyculin A-treated (purple histogram) normal human peripheral blood cells with Anti-Human/Mouse phospho-IkBa (S32/S36) PE. Cells in the lymphocyte gate were used for analysis. Cells were stained using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol.

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