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Catalog No: OKCD01040 (Formerly GWB-KBBOC9)
Size:96WELLS
Price: $700.00
SKU
OKCD01040
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Datasheets/ManualsPrintable datasheet for ITK ELISA Kit (Mouse) (OKCD01040)
Product Info
Predicted Species ReactivityMouse|Mus musculus
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 IL2 Inducible T-Cell Kinase (ITK). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to IL2 Inducible T-Cell Kinase (ITK). 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 IL2 Inducible T-Cell Kinase (ITK), 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 IL2 Inducible T-Cell Kinase (ITK) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
ELISA Kit Range0.312-20ng/mL
ELISA Kit ReproducibilityIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level IL2 Inducible T-Cell Kinase (ITK) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level IL2 Inducible T-Cell Kinase (ITK) 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-Itk Microplate96 Wells (12 x 8 Well Strips)
Itk Lyophilized Standard2 x 20 ng
100X Biotinylated Itk 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< 0.119 ng/mL
SpecificityThis assay has high sensitivity and excellent specificity for detection of IL2 Inducible T-Cell Kinase (ITK). No significant cross-reactivity or interference between IL2 Inducible T-Cell Kinase (ITK) and analogues was observed.
Assay InfoAssay Methodology: Quantitative Sandwich Immunoassay
Gene SymbolItk
Gene Full NameIL2 inducible T cell kinase
Alias SymbolsEmt;IL-2-inducible T-cell kinase;interleukin-2-inducible T cell kinase;Kinase EMT;kinase TLK;T-cell-specific kinase;Tcs;Tcsk;Ts;Tsk;tyrosine-protein kinase ITK/TSK.
NCBI Gene Id16428
Protein NameTyrosine-protein kinase ITK/TSK
Description of TargetTyrosine kinase that plays an essential role in regulation of the adaptive immune response. Regulates the development, function and differentiation of conventional T-cells and nonconventional NKT-cells. When antigen presenting cells (APC) activate T-cell receptor (TCR), a series of phosphorylation lead to the recruitment of ITK to the cell membrane, in the vicinity of the stimulated TCR receptor, where it is phosphorylated by LCK. Phosphorylation leads to ITK autophosphorylation and full activation. Once activated, phosphorylates PLCG1, leading to the activation of this lipase and subsequent cleavage of its substrates. In turn, the endoplasmic reticulum releases calcium in the cytoplasm and the nuclear activator of activated T-cells (NFAT) translocates into the nucleus to perform its transcriptional duty. Phosphorylates 2 essential adapter proteins: the linker for activation of T-cells/LAT protein and LCP2. Then, a large number of signaling molecules such as VAV1 are recruited and ultimately lead to lymphokine production, T-cell proliferation and differentiation. Phosphorylates TBX21 at 'Tyr-525' and mediates its interaction with GATA3 (PubMed:15662016).
Uniprot IDQ03526
Protein Accession #NP_001268894
Nucleotide Accession #NM_001281965.1
Protein Size (# AA)625
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