Human Transcription Factor A, Mitochondrial ELISA Kit

From Antibodies.com, this sandwich ELISA kit is intended for human samples and enables the in vitro quantitative determination of human mtTFA in tissue homogenates, cell lysates, or other biological fluids. The assay uses an antibody specific for mtTFA pre-coated onto a 96-well microplate configured as 12 x 8 well strips.

Standards and test samples are added to the wells, where any mtTFA present is captured by the immobilised antibody. After incubation and washing, a Biotinylated Anti-mtTFA Antibody is added, followed by HRP-Avidin conjugate. A TMB substrate solution is then used to visualise the HRP enzymatic reaction, producing a blue product that turns yellow upon addition of acidic stop solution. The intensity of the yellow colour, read as O.D. absorbance at 450 nm in a colorimetric microplate reader, is proportional to the amount of mtTFA captured, allowing the concentration to be calculated against a standard curve.

The kit has an assay range of 0.312–20 ng/ml and a sensitivity of <0.109 ng/ml, with a total assay time of 4h 30m. Reported recovery in serum (n=5) ranged from 80%–102% (average 91%), in EDTA plasma (n=5) from 81%–100% (average 90%), and in heparin plasma (n=5) from 80%–89% (average 84%). Dilution linearity data for serum, EDTA plasma, and heparin plasma at dilutions of 1:2, 1:4, 1:8, and 1:16 are also supplied. Components should be stored at +4°C or –20°C as instructed.