Human VGF/Nerve Growth Factor Inducible ELISA Kit

A Sandwich ELISA, Double Antibody assay from Assay Genie designed for the quantitative detection of human VGF (Nerve Growth Factor Inducible). The kit is intended for use with serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. It shows a sensitivity of 46.875 pg/mL and a standard range of 78.125-5000 pg/mL. The kit is stored at 2-8°C for 12 months. VGF, also known as neurosecretory protein VGF (Uniprot O15240), is a protein involved in nerve growth factor signaling that promotes neuronal development and function and is relevant to research in neurological disorders, mental health conditions, and metabolic diseases.

Performance characteristics include linearity across dilutions: at 1:2, 1:4 and 1:8, serum (n=5) shows 85-104%, 82-95% and 83-95%; EDTA plasma (n=5) shows 87-100%, 85-96% and 82-94%; heparin plasma (n=5) shows 87-105%, 84-91% and 93-99%. Recovery values are serum (n=5) 86-105% with an average of 94%, EDTA plasma (n=5) 90-104% averaging 97%, and heparin plasma (n=5) 85-100% averaging 93%. The assay procedure is a six-step sandwich ELISA protocol: equilibrate reagents and TMB substrate to room temperature and set up standards, samples and blanks on the pre-coated plate; incubate at 37°C for 90 minutes; add biotin-labeled detection antibody and incubate at 37°C for 60 minutes; add HRP-Streptavidin (SABC) and incubate at 37°C for 30 minutes; add TMB substrate and incubate in the dark for 10-20 minutes; add stop solution and read absorbance at 450 nm immediately.

Sample preparation guidance from the manufacturer includes allowing blood to clot and centrifuging serum at 1000 × g for 20 minutes, collecting plasma via anticoagulant tubes centrifuged 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. Note: The protocol provided is a sample protocol and is specific to each batch/lot; the protocol included with the kit should be followed.