Human Protein inhibitor of activated STAT3 (PIAS3) ELISA Kit

E3 SUMO-protein ligase PIAS3 is a member of the PIAS family of transcriptional modulators. The protein functions as a SUMO (small ubiquitin-like modifier) -E3 ligase which catalyzes the covalent attachment of a SUMO protein to specific target substrates. It directly binds to several transcription factors and either blocks or enhances their activity. Alternatively spliced transcript variants of this gene have been identified, but the full-length nature of some of these variants has not been determined.Gel-shift assays showed that PIAS3 can block MITF DNA-binding activity, and cotransfection of MITF and PIAS3 in NIH 3T3 cells inhibited MITF-driven transcription activity. Interaction between PIAS3 and MITF was independent of STAT3 binding.

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Human Protein inhibitor of activated STAT3 (PIAS3) ELISA Kit has high sensitivity and excellent specificity for detection of Human PIAS3. No significant cross-reactivity or interference between Human PIAS3 and analogues was observed.

This Human Protein inhibitor of activated STAT3 (PIAS3) ELISA Kit employs a two-site sandwich ELISA to quantitate PIAS3 in samples. An antibody specific for PIAS3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPIAS3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PIAS3 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 PIAS3 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.