GM-CSF was initially characterized as a factor that can support the in vitro colony formation of granulocyte-macrophage progenitors. It is also a growth factor for erythroid, megakaryocyte, and eosinophil progenitors. GM-CSF is produced by a number of different cell types (including T cells, B cells, macrophages, mast cells, endothelial cells, fibroblasts, and adipocytes) in response to cytokine or inflammatory stimuli. On mature hematopoietic cells, GM-CSF is a survival factor for and activates the effector functions of granulocytes, monocytes/macrophages, and eosinophils. GM-CSF promotes a Th1 biased immune response, angiogenesis, allergic inflammation, and the development of autoimmunity. It shows clinical effectiveness in ameliorating chemotherapy-induced neutropenia, and GM-CSF transfected tumor cells are utilized as cancer vaccines. The 22kD glycosylated GM-CSF, similar to IL-3 and IL-5, is a cytokine with a core of four bundled alpha helices. Mature human GM-CSF shares 63% - 70%aa sequence identity with canine, feline, porcine, and rat GM-CSF and 54% with mouse GM-CSF. GM-CSF exerts its biological effects through a heterodimeric receptor complex composed of GM-CSF R alpha/CD116 and the signal transducing common beta chain (CD131) which is also a component of the high-affinity receptors for IL-3 and IL-5. In addition, GM-CSF binds a naturally occurring soluble form of GM-CSF R alpha. Human GM-CSF is active on canine and feline cells but not on murine cells.
Sample Type:Cell culture supernatants, serum, plasma, and tissue
Intended Use:The Human GM-CSF BioAssay™ ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human GM-CSF concentrations in cell culture supernatants, serum, plasma, and tissue.
Sensitivity:4pg/ml
Range:15.625-1000pg/ml
Specificity:Recognizes human GM-CSF.
Sample Volume:100ul/well
Test Principle:This assay employs the Sandwich immunoassay technique. An anti-h GM-CS Fmonoclonal coating antibody is adsorbed onto microwells. GM-CSF present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-h GM-CSF antibody is added and binds to GM-CSF captured by the first antibody. Following incubation unbound Biotinylated anti-h GM-CSF antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h GM-CSF antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of GM-CSF present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven GM-CSF standard dilutions and GM-CSF sample concentration determined.
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
