Human MED30 (Mediator complex subunit 30) ELISA Kit (HUFI07642)

A colorimetric Sandwich ELISA Kit from Assay Genie is offered for the quantitative measurement of human MED30 (Mediator complex subunit 30). The double-antibody sandwich format is rated with a sensitivity of 46.875 pg/mL and a working range of 78.125–5000 pg/mL, and is validated for use with serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. Reagents are supplied for storage at 2–8°C for 12 months.

The six-step workflow comprises: equilibrating reagents and the pre-coated plate to room temperature; loading standards, samples and blanks and incubating at 37°C for 90 minutes; adding biotin-labelled detection antibody and incubating at 37°C for 60 minutes; adding HRP-Streptavidin (SABC) and incubating at 37°C for 30 minutes; developing with TMB substrate in the dark for 10–20 minutes; and adding stop solution before reading absorbance at 450 nm. Sample-handling guidance covers serum (clot, centrifuge at 1000 × g for 20 minutes), plasma (anticoagulant tubes, centrifuge at 1000 × g for 15 minutes at 2–8°C), tissue homogenate, cell culture supernatant (centrifuge at 2500 rpm for 5 minutes) and cell lysate preparations.

Reported dilution linearity at 1:2, 1:4 and 1:8 is 89–102%, 85–95% and 92–98% for serum (n=5); 93–105%, 86–97% and 91–100% for EDTA plasma (n=5); and 89–105%, 87–101% and 92–97% for heparin plasma (n=5). Recovery ranges with average values are: serum (n=5) 87–94% averaging 97%; EDTA plasma (n=5) 85–104% averaging 90%; heparin plasma (n=5) 88–98% averaging 94%. MED30 is described as a subunit of the Mediator complex that contributes to transcriptional regulation and gene expression.