IKBKB ELISA Kit (Mouse)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Inhibitor Of Kappa-Light Polypeptide Gene Enhancer In B-Cells Kinase Beta (IkBKb) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Inhibitor Of Kappa-Light Polypeptide Gene Enhancer In B-Cells Kinase Beta (IkBKb) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

AI132552; I-kappa-B kinase 2; I-kappa-B-kinase beta; IKK; IKK-; IKK[b; IKK[b]; IKK2; IKK-2; IKK-B; IKKbe; IKK-be; IKKbeta; IKK-beta; inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta; inhibitor of nuclear factor kappa-B kinase subunit beta; NFKBIKB; nuclear factor NF-kappa-B inhibitor kinase beta; serine/threonine protein kinase IKBKB.

Serine kinase that plays an essential role in the NF-kappa-B signaling pathway which is activated by multiple stimuli such as inflammatory cytokines, bacterial or viral products, DNA damages or other cellular stresses. Acts as part of the canonical IKK complex in the conventional pathway of NF-kappa-B activation and phosphorylates inhibitors of NF-kappa-B on 2 critical serine residues. These modifications allow polyubiquitination of the inhibitors and subsequent degradation by the proteasome. In turn, free NF-kappa-B is translocated into the nucleus and activates the transcription of hundreds of genes involved in immune response, growth control, or protection against apoptosis. In addition to the NF-kappa-B inhibitors, phosphorylates several other components of the signaling pathway including NEMO/IKBKG, NF-kappa-B subunits RELA and NFKB1, as well as IKK-related kinases TBK1 and IKBKE. IKK-related kinase phosphorylations may prevent the overproduction of inflammatory mediators since they exert a negative regulation on canonical IKKs. Phosphorylates FOXO3, mediating the TNF-dependent inactivation of this pro-apoptotic transcription factor. Also phosphorylates other substrates including NCOA3, BCL10 and IRS1. Within the nucleus, acts as an adapter protein for NFKBIA degradation in UV-induced NF-kappa-B activation.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Inhibitor Of Kappa-Light Polypeptide Gene Enhancer In B-Cells Kinase Beta (IkBKb) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Inhibitor Of Kappa-Light Polypeptide Gene Enhancer In B-Cells Kinase Beta (IkBKb) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Inhibitor Of Kappa-Light Polypeptide Gene Enhancer In B-Cells Kinase Beta (IkBKb), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Inhibitor Of Kappa-Light Polypeptide Gene Enhancer In B-Cells Kinase Beta (IkBKb) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery range (%) Average (%) Serum (n=5) 82-99 94 EDTA Plasma (n=5) 79-93 86 Heparin Plasma (n=5) 87-101 93 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 82-99 94 Serum (n=5) 82-99 94 EDTA Plasma (n=5) 79-93 86 EDTA Plasma (n=5) 79-93 86 Heparin Plasma (n=5) 87-101 93 Heparin Plasma (n=5) 87-101 93

Component Amount Anti-IKBKB Microplate 96 Wells (12 x 8 Well strips) IKBKB Lyophilized Standard 2 100X Biotinylated IKBKB Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-IKBKB Microplate 96 Wells (12 x 8 Well strips) Anti-IKBKB Microplate 96 Wells (12 x 8 Well strips) IKBKB Lyophilized Standard 2 IKBKB Lyophilized Standard 2 100X Biotinylated IKBKB Detector Antibody 1 x 120 uL 100X Biotinylated IKBKB Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

This assay has high sensitivity and excellent specificity for detection of Inhibitor Of Kappa-Light Polypeptide Gene Enhancer In B-Cells Kinase Beta (IkBKb) . No significant cross-reactivity or interference between Inhibitor Of Kappa-Light Polypeptide Gene Enhancer In B-Cells Kinase Beta (IkBKb) and analogues was observed.