Human Connexin 32/GJB1 ELISA Kit

From Assay Genie, this Sandwich ELISA kit enables quantitative measurement of human Connexin 32 (GJB1) by colorimetric detection. The assay is validated for serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples, supporting research into gap junction biology. Connexin 32, also referred to as GJB1, is a gap junction protein whose dysregulation has been associated with Charcot-Marie-Tooth disease, a hereditary peripheral neuropathy.

Performance characteristics include a sensitivity of 0.188 ng/mL and a standard curve range of 0.313-20 ng/mL. Linearity across 1:2, 1:4, and 1:8 dilutions was 85-104%, 85-105%, and 86-102% for serum (n = 5); 82-99%, 82-100%, and 82-87% for EDTA plasma (n = 5); and 82-97%, 85-100%, and 80-99% for heparin plasma (n = 5). Recovery ranges and averages were 88-100% (94%) for serum, 88-100% (94%) for EDTA plasma, and 88-102% (97%) for heparin plasma. Unopened reagents are stored at 2-8°C for 12 months.

The workflow begins with reagent equilibration, followed by a 90-minute primary incubation at 37°C, a 60-minute biotin-labeled detection antibody incubation at 37°C, a 30-minute HRP-Streptavidin (SABC) binding step at 37°C, TMB color development in the dark for 10-20 minutes, and absorbance reading at 450 nm after stop solution addition. Serum is processed by clotting followed by centrifugation at 1000 × g for 20 minutes; plasma is collected into anticoagulant tubes and centrifuged at 1000 × g for 15 minutes at 2-8°C.