Mouse TRAF2 and NCK-interacting protein kinase (TNIK) ELISA Kit

TNIK was autophosphorylated in a manner dependent upon lys54 in the ATP-binding pocket of its kinase domain. Immunoprecipitation analysis showed that epitope-tagged TNIK coprecipitated endogenous TRAF2 from human embryonic kidney cells. Mutation analysis revealed that the intermediate domain was sufficient for the interaction, although the GCK domain may contribute. The intermediate domain was also sufficient for interaction with NCK. Cotransfection of TNIK with JNK2 (MAPK9) enhanced JNK2 kinase activity in a dose-dependent manner, and this effect was mediated by the GCK region of TNIK, but not the kinase domain. TNIK overexpression had no effect on ERK1 (MAPK3), p38 (MAPK14), or NF-kappa-B.

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Mouse TRAF2 and NCK-interacting protein kinase (TNIK) ELISA Kit has high sensitivity and excellent specificity for detection of Mouse TNIK. No significant cross-reactivity or interference between Mouse TNIK and analogues was observed.

This Mouse TRAF2 and NCK-interacting protein kinase (TNIK) ELISA Kit employs a two-site sandwich ELISA to quantitate TNIK in samples. An antibody specific for TNIK has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyTNIK present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for TNIK is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of TNIK bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.