Phospho-IkB alpha (Ser32, Ser36) Monoclonal Antibody (RILYB3R), eFluor™ 660, eBioscience™, Invitrogen™

Description

• 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
• 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.