BRCC3 ELISA Kit (Human)

ELISA Kit

Root Catalog/Products/ELISA Kits, Root Catalog/Products/Kit

BRCC3

316

Metalloprotease that specifically cleaves 'Lys-63'-linked polyubiquitin chains (PubMed:19214193, PubMed:20656690, PubMed:24075985, PubMed:26344097). Does not have activity toward 'Lys-48'-linked polyubiquitin chains. Component of the BRCA1-A complex, a complex that specifically recognizes 'Lys-63'-linked ubiquitinated histones H2A and H2AX at DNA lesions sites, leading to target the BRCA1-BARD1 heterodimer to sites of DNA damage at double-strand breaks (DSBs). In the BRCA1-A complex, it specifically removes 'Lys-63'-linked ubiquitin on histones H2A and H2AX, antagonizing the RNF8-dependent ubiquitination at double-strand breaks (DSBs) (PubMed:20656690). Catalytic subunit of the BRISC complex, a multiprotein complex that specifically cleaves 'Lys-63'-linked ubiquitin in various substrates (PubMed:20656690, PubMed:24075985, PubMed:26344097, PubMed:26195665). Mediates the specific 'Lys-63'-specific deubiquitination associated with the COP9 signalosome complex (CSN), via the interaction of the BRISC complex with the CSN complex (PubMed:19214193). The BRISC complex is required for normal mitotic spindle assembly and microtubule attachment to kinetochores via its role in deubiquitinating NUMA1 (PubMed:26195665). Plays a role in interferon signaling via its role in the deubiquitination of the interferon receptor IFNAR1; deubiquitination increases IFNAR1 activity by enhancing its stability and cell surface expression (PubMed:24075985, PubMed:26344097). Down-regulates the response to bacterial lipopolysaccharide (LPS) via its role in IFNAR1 deubiquitination (PubMed:24075985).

NP_001018065

This assay has high sensitivity and excellent specificity for detection of BRCA1/BRCA2 Containing Complex Subunit 3 (BRCC3).No significant cross-reactivity or interference between BRCA1/BRCA2 Containing Complex Subunit 3 (BRCC3) and analogues was observed.

Component: Amount
Anti-BRCC3 Microplate: 96 Wells (12 x 8 Well Strips)
BRCC3 Lyophilized Standard: 2 x 10 ng
100X Biotinylated BRCC3 Detector Antibody: 120 uL
Avidin/HRP Conjugate: 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
TMB Substrate: 1 x 9 mL
Stop Solution: 1 x 6 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 BRCA1/BRCA2 Containing Complex Subunit 3 (BRCC3). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to BRCA1/BRCA2 Containing Complex Subunit 3 (BRCC3). 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 BRCA1/BRCA2 Containing Complex Subunit 3 (BRCC3), 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 BRCA1/BRCA2 Containing Complex Subunit 3 (BRCC3) 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 BRCA1/BRCA2 Containing Complex Subunit 3 (BRCC3) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level BRCA1/BRCA2 Containing Complex Subunit 3 (BRCC3) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%