Description
Principle of the Assay::The Human FGF2 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human FGF2 in Cell Culture Supernatants, Serum, Plasma, Tissue Homogenates. This assay employs an antibody specific for Human FGF2 coated on a 96-well plate. Standards and samples are pipetted into the wells and FGF2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human FGF2 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of FGF2 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Background/Introduction: Human FGF-Basic or Fibroblast Growth Factor 2, also known commonly as HBGF-2 or Heparin-Binding Growth Factor 2, is a 288 amino acid cytokine protein encoded by the FGF2gene at locus 4q26 on chromosome 4. FGF family members bind heparin and possess broad mitogenic and angiogenic activities. This protein has been implicated in diverse biological processes, such as limb and nervous system development, would healing and tumor growth. The mRNA for this gene contains multiple polyadenylation sites, and is alternatively translated from non-AUG (CUG) and AUG initiation codons, resulting in five different isoforms with distinct properties. The CUG-initiated isoforms are localized in the nucleus and are responsible for the intracrine effect, whereas, the AUG-initiated form is mostly cytosolic and is responsible for the paracrine and autocrine effects of this FGF. The monomeric FGF-Basic protein interacts with many other proteins such as CSPG4 and FGFBP1 while also believed to be found in a complex with FGFBP1, FGF1 and FGF2. Expression patterns for this protein typically take place in granulosa and cumulus cells, but FGF2 is also found in hepatocellular carcinoma cells (not non-cancerous liver tissue). Several experiments were performed on FGF-Basic to determine its pH and temperature properties. It has been recently determined that FGF-Basic retains almost half of its activity after treatment at pH 2.0 for 3 hours at 20 degree C, while being inactivated after 3 minutes at 60 degree C or 1 minute at 80 degree C