Seprase/FAP (FAP) ELISA Kit for Human

Background: FAP (Fibroblast Activation Protein, Alpha) also known as FAPA or SEPRASE, is an inducible cell surface glycoprotein that was originally identified in cultured fibroblasts using monoclonal antibody F19. The protein encoded by this gene is a homodimeric integral membrane gelatinase belonging to the serine protease family. The FAP gene is mapped on 2q24.2. FAP is most closely related to DPPIV and they share about 50% of their amino acids. FAP is catalytically active as a 170kD dimer and has dipeptidase and gelatinase activity. Its gelatinase activity requires a glycine in P2 position.FAPalpha shows 48% amino acid identity with dipeptidyl peptidase IV and 30% identity with DPP4-related protein. Northern blot analysis detected a 2.8-kb FAP-alpha mRNA in fibroblasts. Depletion of FAP-expressing cells, which made up only 2% of all tumor cells in established Lewis lung carcinomas, caused rapid hypoxic necrosis of both cancer and stromal cells in immunogenic tumors by a process involving interferon-gamma and tumor necrosis factor-alpha.

Principle of the Assay: The Picokine™ Human FAP Pre-Coated ELISA (Enzyme-Linked Immunosorbent Assay) kit is a solid-phase immunoassay specially designed to measure Human FAP with a 96-well strip plate that is pre-coated with antibody specific for FAP. The detection antibody is a biotinylated antibody specific for FAP. The capture antibody is monoclonal antibody from mouse and the detection antibody is polyclonal antibody from goat. The kit includes Human FAP protein as standards. To measure Human FAP, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS or TBS buffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS or TBS buffer and add TMB. TMB is an HRP substrate and will be catalyzed to produce a blue color product, which changes into yellow after adding the acidic stop solution. The absorbance of the yellow product at 450nm is linearly proportional to Human FAP in the sample. Read the absorbance of the yellow product in each well using a plate reader, and benchmark the sample wells' readings against the standard curve to determine the concentration of Human FAP in the sample.

Small volumes of FAP elisa kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS177274 is a ready-to-use microwell, strip plate ELISA (enzyme-linked immunosorbent assay) Kit for analyzing the presence of the Seprase/FAP, ELISA Kit target analytes in biological samples. The concentration gradients of the kit standards or positive controls render a theoretical kit detection range in biological research samples containing FAP. The ELISA analytical biochemical technique of the MBS177274 kit is based on FAP antibody-FAP antigen interactions (immunosorbency) and an HRP colorimetric detection system to detect FAP antigen targets in samples. The ELISA Kit is designed to detect native, not recombinant, FAP. Appropriate sample types may include undiluted body fluids and/or tissue homogenates, secretions. Quality control assays assessing reproducibility identified the intra-assay CV (%) and inter-assay CV(%).