Description
Principle of the assay: rat GM-CSF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for GM-CSF has been precoated onto 96-well plates. Standards (E.coli, A1-K127) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for GM-CSF is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the rat GM-CSF amount of sample captured in plate.
Background: Granulocyte-macrophage colony-stimulating factor (GM-CSF) is also symbolized CSF2. Human GM-CSF is a glycoprotein that is essential for the in vitro proliferation and differentiation of precursor cells into mature granulocytes and macrophages. The human cDNA clones contain a single open-reading frame encoding a protein of 144 amino acids with a predicted molecular mass of 16,293 daltons and show 69% nucleotide homology and 54% amino acid homology to mouse GM-CSF. The gene for human GM-CSF appears to exist as a single-copy gene.1 Human GM-CSF is a 22,000-dalton glycoprotein that stimulates the growth of myeloid progenitor cells andacts directly on mature neutrophils. The GM-CSF gene is localized by somatic cell hybrid analysis and in situ hybridization to human chromosome region 5q21-5q32, which is involved in interstitial deletions in the 5q- syndrome and acute myelogenous leukemia.2 A complementary DNA for the T lymphocyte-derived lymphokine,GM-CSF has been cloned, and recombinant GM-CSF protein has been expressed in yeast and purified to homogeneity. This purified human recombinant GM-CSF stimulates peripheral blood monocytes in vitro to become cytotoxic for the malignant melanoma cell line A375.3