PIAS3 ELISA Kit (Human)

ELISA Kit

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PIAS3

628

Functions as an E3-type small ubiquitin-like modifier (SUMO) ligase, stabilizing the interaction between UBE2I and the substrate, and as a SUMO-tethering factor. Plays a crucial role as a transcriptional coregulation in various cellular pathways, including the STAT pathway and the steroid hormone signaling pathway. Involved in regulating STAT3 signaling via inhibiting STAT3 DNA-binding and suppressing cell growth. Enhances the sumoylation of MTA1 and may participate in its paralog-selective sumoylation (PubMed:21965678, PubMed:9388184). Sumoylates CCAR2 which promotes its interaction with SIRT1 (PubMed:25406032). Diminishes the sumoylation of ZFHX3 by preventing the colocalization of ZFHX3 with SUMO1 in the nucleus (PubMed:24651376).

NP_006090.2

This assay has high sensitivity and excellent specificity for detection of Protein Inhibitor Of Activated STAT 3 (PIAS3).No significant cross-reactivity or interference between Protein Inhibitor Of Activated STAT 3 (PIAS3) and analogues was observed.

Component: Amount
Anti-PIAS3 Microplate: 96 Wells (12 x 8 Well strips)
PIAS3 Lyophilized Standard: 2
100X Biotinylated PIAS3 Detector Antibody: 1 x 120 uL
100X Avidin-HRP Conjugate: 1 x 120 uL
Standard Diluent: 1 x 20 mL
Detector Antibody Diluent: 1 x 12 mL
Conjugate Diluent: 1 x 12 mL
30X Wash Buffer: 1 x 20 mL
Stop Solution: 1 x 6 mL
TMB Substrate: 1 x 9 mL

3h

The 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 Protein Inhibitor Of Activated STAT 3 (PIAS3). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Protein Inhibitor Of Activated STAT 3 (PIAS3). 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 Protein Inhibitor Of Activated STAT 3 (PIAS3), 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 Protein Inhibitor Of Activated STAT 3 (PIAS3) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Protein Inhibitor Of Activated STAT 3 (PIAS3) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Protein Inhibitor Of Activated STAT 3 (PIAS3) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%