CHUK ELISA Kit (Bovine)

Conserved helix-loop-helix ubiquitous kinase; i kappa-B kinase alpha; I-kappa-B kinase 1; I-kappa-B kinase alpha; ikBKA; IKK1; IKK-A; IKK-alpha; inhibitor of nuclear factor kappa-B kinase subunit alpha; NFKBIKA; nuclear factor NF-kappa-B inhibitor kinase alpha.

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 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. Negatively regulates the pathway by phosphorylating the scaffold protein TAXBP1 and thus promoting the assembly of the A20/TNFAIP3 ubiquitin-editing complex (composed of A20/TNFAIP3, TAX1BP1, and the E3 ligases ITCH and RNF11) . Therefore, CHUK plays a key role in the negative feedback of NF-kappa-B canonical signaling to limit inflammatory gene activation. As part of the non-canonical pathway of NF-kappa-B activation, the MAP3K14-activated CHUK/IKKA homodimer phosphorylates NFKB2/p100 associated with RelB, inducing its proteolytic processing to NFKB2/p52 and the formation of NF-kappa-B RelB-p52 complexes. In turn, these complexes regulate genes encoding molecules involved in B-cell survival and lymphoid organogenesis. Participates also in the negative feedback of the non-canonical NF-kappa-B signaling pathway by phosphorylating and destabilizing MAP3K14/NIK. Within the nucleus, phosphorylates CREBBP and consequently increases both its transcriptional and histone acetyltransferase activities. Modulates chromatin accessibility at NF-kappa-B-responsive promoters by phosphorylating histones H3 at 'Ser-10' that are subsequently acetylated at 'Lys-14' by CREBBP. Additionally, phosphorylates the CREBBP-interacting protein NCOA3. Also phosphorylates FOXO3 and may regulate this pro-apoptotic transcription factor.

Aviva Systems Biology Chuk ELISA Kit (Bovine) (OKWB00111) is based on standard sandwich enzyme-linked immuno-sorbent assay technology. An antibody specific for Chuk has been pre-coated onto a 96-well plate (12 x 8 Well Strips) . Standards or test samples are added to the wells, incubated and removed. A biotinylated detector antibody specific for Chuk is added, incubated and followed by washing. Avidin-Peroxidase Conjugate is then added, incubated and unbound conjugate is washed away. An enzymatic reaction is produced through the addition of TMB substrate which is catalyzed by HRP generating a blue color product that changes to yellow after adding acidic stop solution. The density of yellow coloration read by absorbance at 450 nm is quantitatively proportional to the amount of sample Chuk captured in the well.

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

Sample Type Range Mean Serum (n=5) 89-103 97 EDTA Plasma (n=5) 94-103 99 Heparin Plasma (n=5) 86-102 94 Sample Type Range Mean Sample Type Range Mean Serum (n=5) 89-103 97 Serum (n=5) 89-103 97 EDTA Plasma (n=5) 94-103 99 EDTA Plasma (n=5) 94-103 99 Heparin Plasma (n=5) 86-102 94 Heparin Plasma (n=5) 86-102 94

Component Amount Chuk Microplate 96 Wells (12 x 8 Well strips) Chuk Lyophilized Standard 2 x 10 ng 100X Chuk Biotinylated Detector Anitbody 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 25X Wash Buffer 1 x 20 mL TMB Substrate 1 x 10 mL Stop Solution 1 x 10 mL Component Amount Component Amount Chuk Microplate 96 Wells (12 x 8 Well strips) Chuk Microplate 96 Wells (12 x 8 Well strips) Chuk Lyophilized Standard 2 x 10 ng Chuk Lyophilized Standard 2 x 10 ng 100X Chuk Biotinylated Detector Anitbody 1 x 120 uL 100X Chuk Biotinylated Detector Anitbody 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 25X Wash Buffer 1 x 20 mL 25X Wash Buffer 1 x 20 mL TMB Substrate 1 x 10 mL TMB Substrate 1 x 10 mL Stop Solution 1 x 10 mL Stop Solution 1 x 10 mL