Human Serine/threonine-protein phosphatase PGAM5, mitochondrial (PGAM5) ELISA Kit

Keap1 is a BTB-Kelch substrate adaptor protein for a Cul3-dependent ubiquitin ligase complex that functions as a sensor for thiol-reactive chemopreventive compounds and oxidative stress. PGAM5, as a novel substrate for Keap1. The N terminus of the PGAM5 protein contains a conserved NXESGE motif that binds to the substrate binding pocket in the Kelch domain of Keap1, whereas the C-terminal PGAM domain binds Bcl-XL. Keap1-dependent ubiquitination of PGAM5 results in proteasome-dependent degradation of PGAM5. Quinone-induced oxidative stress and the chemopreventive agent sulforaphane inhibit Keap1-dependent ubiquitination of PGAM5. The identification of PGAM5 as a novel substrate of Keap1 suggests that Keap1 regulates both transcriptional and post-transcriptional responses of mammalian cells to oxidative stress.

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Human Serine/threonine-protein phosphatase PGAM5, mitochondrial (PGAM5) ELISA Kit has high sensitivity and excellent specificity for detection of Human PGAM5. No significant cross-reactivity or interference between Human PGAM5 and analogues was observed.

This Human Serine/threonine-protein phosphatase PGAM5, mitochondrial (PGAM5) ELISA Kit employs a two-site sandwich ELISA to quantitate PGAM5 in samples. An antibody specific for PGAM5 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPGAM5 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PGAM5 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 PGAM5 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.