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Catalog No: OKCD00674 (Formerly GWB-KBBGH3)
Size:96WELLS
Price: $650.00
SKU
OKCD00674
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Datasheets/ManualsPrintable datasheet for IRAK4 ELISA Kit (Human) (OKCD00674)
Product Info
Predicted Species ReactivityHomo sapiens|Human
ApplicationEnzyme-linked Immunosorbent assay-Sandwich
ELISA Kit Detection MethodColorimetric
ELISA Kit Duration3h
ELISA Kit PrincipleThe 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 Interleukin 1 Receptor Associated Kinase 4 (IRAK4). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Interleukin 1 Receptor Associated Kinase 4 (IRAK4). 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 Interleukin 1 Receptor Associated Kinase 4 (IRAK4), 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 Interleukin 1 Receptor Associated Kinase 4 (IRAK4) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
ELISA Kit Range1.56-100ng/mL
ELISA Kit ReproducibilityIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Interleukin 1 Receptor Associated Kinase 4 (IRAK4) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Interleukin 1 Receptor Associated Kinase 4 (IRAK4) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
ELISA Kit Component
ComponentAmount
Anti-IRAK4 Microplate96 Wells (12 x 8 Well Strips)
IRAK4 Lyophilized Standard2 x 100 ng
100X Biotinylated IRAK4 Detector Antibody120 uL
Avidin/HRP Conjugate120 uL
Standard Diluent1 x 20 mL
Detector Antibody Diluent1 x 12 mL
Conjugate Diluent1 x 12 mL
30X Wash Buffer1 x 20 mL
TMB Substrate1 x 9 mL
Stop Solution1 x 6 mL
Reconstitution and Storage2°C to 8°C|-20°C
Sample TypeTissue homogenates, cell lysates and other biological fluids
Sensitivity< 0.64 ng/mL
SpecificityThis assay has high sensitivity and excellent specificity for detection of Interleukin 1 Receptor Associated Kinase 4 (IRAK4). No significant cross-reactivity or interference between Interleukin 1 Receptor Associated Kinase 4 (IRAK4) and analogues was observed.
Assay InfoAssay Methodology: Quantitative Sandwich Immunoassay
Gene SymbolIRAK4
Gene Full Nameinterleukin 1 receptor associated kinase 4
Alias SymbolsIMD67;interleukin-1 receptor-associated kinase 4;IPD1;IRAK-4;NY-REN-64;REN64;renal carcinoma antigen NY-REN-64.
NCBI Gene Id51135
Protein NameInterleukin-1 receptor-associated kinase 4
Description of TargetSerine/threonine-protein kinase that plays a critical role in initiating innate immune response against foreign pathogens. Involved in Toll-like receptor (TLR) and IL-1R signaling pathways (PubMed:17878374). Is rapidly recruited by MYD88 to the receptor-signaling complex upon TLR activation to form the Myddosome together with IRAK2. Phosphorylates initially IRAK1, thus stimulating the kinase activity and intensive autophosphorylation of IRAK1. Phosphorylates E3 ubiquitin ligases Pellino proteins (PELI1, PELI2 and PELI3) to promote pellino-mediated polyubiquitination of IRAK1. Then, the ubiquitin-binding domain of IKBKG/NEMO binds to polyubiquitinated IRAK1 bringing together the IRAK1-MAP3K7/TAK1-TRAF6 complex and the NEMO-IKKA-IKKB complex. In turn, MAP3K7/TAK1 activates IKKs (CHUK/IKKA and IKBKB/IKKB) leading to NF-kappa-B nuclear translocation and activation. Alternatively, phosphorylates TIRAP to promote its ubiquitination and subsequent degradation. Phosphorylates NCF1 and regulates NADPH oxidase activation after LPS stimulation suggesting a similar mechanism during microbial infections.
Uniprot IDQ9NWZ3
Protein Accession #NP_001107654
Nucleotide Accession #NM_001114182.2
Protein Size (# AA)460
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