Human Insulin Like Growth Factor Binding Protein 3 ELISA Kit

Manufactured for Antibodies.com, this sandwich ELISA kit enables the in vitro quantitative determination of human Insulin Like Growth Factor Binding Protein 3 (IGFBP3) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. An antibody specific for IGFBP3 is pre-coated onto a 96-well microplate; standards and samples are added and any IGFBP3 present is captured by the immobilised antibody. After washing, a Biotinylated Anti-IGFBP3 Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate is then used to visualise the HRP enzymatic reaction, producing a blue colour that turns yellow on addition of acidic stop solution. The density of yellow is proportional to the amount of IGFBP3 captured, and concentration is calculated from O.D. absorbance at 450 nm referenced to a standard curve.

The standard curve range is 0.156–10 ng/ml with a sensitivity of <0.069 ng/ml, and the total assay time is 4 h 30 min. Detection is performed on a colourimetric microplate reader. Kit components should be stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.

Validation data for n=5 samples per matrix show recovery of 80%–102% (average 91%) in serum, 81%–101% (average 91%) in EDTA plasma and 80%–89% (average 84%) in heparin plasma. Dilution linearity across 1:2, 1:4, 1:8 and 1:16 was 87%–91%, 87%–107%, 74%–101% and 92%–97% in serum; 90%–105%, 84%–101%, 90%–101% and 79%–108% in EDTA plasma; and 84%–95%, 92%–105%, 82%–105% and 89%–91% in heparin plasma.