Monkey Fetuin A/AHSG ELISA Kit

From Assay Genie, this Sandwich ELISA is designed for the precise measurement of monkey fetuin-A (AHSG) levels in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples, using a colorimetric detection method. Fetuin-A (AHSG) is described as a critical glycoprotein involved in regulating insulin sensitivity, inflammation, and bone metabolism, with its dysregulation linked to diabetes, cardiovascular diseases, and liver disorders, making it a useful biomarker for related research.

The double-antibody sandwich assay reports a sensitivity of 3.75 ng/mL and a working range of 6.25-400 ng/mL, with storage at 2-8°C for 12 months. The protocol proceeds by equilibrating reagents and TMB substrate to room temperature, incubating standards and samples at 37°C for 90 minutes, adding a biotin-labelled detection antibody for 60 minutes at 37°C, binding HRP-streptavidin for 30 minutes at 37°C, developing colour with TMB substrate in the dark for 10-20 minutes, and reading absorbance at 450 nm after adding stop solution.

Sample preparation includes allowing blood to clot and centrifuging serum at 1000 × g for 20 minutes, collecting plasma with anticoagulant tubes and centrifuging at 1000 × g for 15 minutes at 2-8°C, homogenising tissue in PBS with protease inhibitors, centrifuging cell culture supernatant at 2500 rpm for 5 minutes, and lysing cells with lysis buffer containing protease inhibitors. Reported linearity spans dilutions of 1:2, 1:4 and 1:8 across serum, EDTA plasma and heparin plasma, with corresponding recovery averages of 95%, 96% and 95%.