IRAK Antibody

IRAK Antibody: Nuclear factor kappa B (NF-κ B) is a ubiquitous transcription factor and an essential mediator of gene expression during activation of immune and inflammatory responses. NF-κ B mediates the expression of a great variety of genes in response to extracellular stimuli including IL-1, TNFα and LPS. A serine/threonine protein kinase associated with IL-1 receptor (IRAK) and its homologue mouse pelle-like protein kinase (mPLK) were identified recently. IRAK is associated with the IL-1 receptor subunits IL-1RI and IL-1RAcP after IL-1 binding and serves as a signaling molecule to mediate IL-1 response. IRAK mediates a signaling cascade leading to NF-κ B activation by members in IL-1 family including IL-1 and a novel cytokine IL-18 (also termed IGIF) .

Independent Antibody Validation in Cell lines (Figure 2) shows similar IRAK expression profile in human cell lines detected by two independent anti-IRAK antibodies that recognize different epitopes, 1007 against C-terminus domain and 64-231 against another region of C-terminus domain. IRAK proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies. Independent Antibody Validation in Cell lines1007 Recombinant Protein Test (Figure 3) : Anti-IRAK antibodies (1007) detected human IRAK recombinant protein at different concentrations. Recombinant Protein Test1007 Immunoprecipitation validation (Figure 6) : IRAK1 protein and pellino protein were immunoprecipitated by anti-IRAK antibodies (1007) in HEK293T cells with co-expression of Pellino proteins and IRAK-1. Immunoprecipitation validation1007 Overexpression validation (Figure 7) : IRAK1 overexpression in 293T cells was detected by anit-IRAK antibodies (1007) . Overexpression validation1007 KD validation (Figure 8) : Anti-IRAK antibody (1007) specificity was further verified by IRAK specific knockdown. IRAK signal in human chondrocytes transfected with IRAK siRNAs was disrupted in comparison with that in cells transfected with control siRNAs. KD validation1007

IRAK antibody can be stored at 4˚ C for three months and -20˚ C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

At least four isoforms of IRAK are known to exist; this antibody will detect all four isoforms. IRAK antibody is predicted to not cross-react with other members of the IRAK protein family.

Human IRAK has 4 isoforms, including isoform 1 (712aa, 77kD), isoform 2 (682aa, 73kD), isoform 3 (693aa, 75kD), and isoform 4 (633aa, 68kD) . Mouse IRAK has 2 isoforms, including isoform 1 (710aa, 77kD) and isoform 2 (712aa, 78kD) . Rat IRAK also has one isoform (710aa, 78kD) . 1007 can detect all human isoforms, mouse and rat IRAK.