Human Apoptosis signal regulating kinase 1 (ASK-1) ELISA Kit

Apoptosis signal-regulating kinase 1 is an enzyme which in humans is encoded by the MAP3K5 gene. This gene is located on chromosome 6 at locus 6q22.33 and the transcribed protein contains 1,374 amino acids with 11 kinase subdomains. The ASK1 (MAPKKK5) protein phosphorylates and activates MKK4 (aliases SERK1, MAPKK4) in vitro, and activates c-Jun N-terminal kinase (JNK) /stress-activated protein kinase (SAPK) during transient expression in COS and 293 cells; ASK1 does not activate MAPK/ERK.ASK1 found in the inactive form, bound to reduced thioredoxin.When oxidized by a reactive oxygen species, Trx dissociates from ASK1. The ASK1, which is found as a homo-oligodimer, autophosphorylates and becomes an active MAP kinase kinase kinase.

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Human Apoptosis signal regulating kinase 1 (ASK-1) ELISA Kit has high sensitivity and excellent specificity for detection of Human MAP3K5. No significant cross-reactivity or interference between Human MAP3K5 and analogues was observed.

This Human Apoptosis signal regulating kinase 1 (ASK-1) ELISA Kit employs a two-site sandwich ELISA to quantitate MAP3K5 in samples. An antibody specific for MAP3K5 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyMAP3K5 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for MAP3K5 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 MAP3K5 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.