Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for GM-CSF has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any GM-CSF present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for GM-CSF is added to the wells and binds to the combination of capture antibody- GM-CSF in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. 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.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Granulocyte macrophage colony stimulating factor (GM-CSF) is a pleiotropic cytokine with multiple effects on hematopoietic cells. It mobilizes CD34+ progenitor cells into the periphery and stimulates their proliferation, survival and differentiation into neutrophils, monocytes/ macrophages, eosinophils, and myeloid dendritic cells. On these terminally differentiated myeloid cells, GM-CSF is also needed for inducing their effector functions. In addition, GM-CSF has been shown to stimulate the proliferation and differentiation of the erythroid and megakaryocyte progenitor cells.
GM-CSF is produced by a number of different cell types, including keratinocytes, mature and immature NK cells, type II alveolar cells), endothelial cells, monocytes, bone-marrow mesenchymal stem cells, CD4+ and CD8+ T cells, megakaryocytes, B cells, eosinophils, chondrocytes and fibroblasts.
Human GM-CSF cDNA encodes a 144 amino acid (aa) residue precursor protein with a 17 aa putative signal peptide and a 127 aa mature protein. Natural GM-CSF is a monomer that contains both N- and O-linked glycosylation. Mature human GM-CSF shares approximately 55%, 63% and 68% aa sequence homology with mouse, rat and canine. GM-CSF, respectively. Human GM-CSF is not biologically active on mouse cells, but was reported to have some activity on canine cells.
GM-CSF exerts its activity through binding to a high affinity receptor complex consisting of two membrane glycoproteins. The presence of the spliced variants in the heteromeric receptor complex can regulate the functions of the complex.
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human GM-CSF (8﹡12)||1 plate|
|Human GM-CSF Standard lyophilized, 2000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-human GM-CSF monoclonal antibody||2 vials|
|Streptavidin-HRP solution||2 vials|
|Standard /sample Diluent||1 bottle|
|Biotin-Conjugate antibody Diluent||1 bottle|
|Streptavidin-HRP Diluent||1 bottle|
|Wash Buffer Concentrate 20x (PBS with 1% Tween-20)||1 bottle|
|Substrate Solution||1 vial|
|Stop Solution||1 vial|
|Adhesive Films||4 pieces|
|Product data sheet||1 copy|
|Storage||Store at 2 - 8°C|
|REPEATABILITY||The coefficient of variation of both intra-assay and inter-assay were less than 10%.|
|SENSITIVITY||The minimum detectable dose was 4pg/mL.|
|ASSAY RANGE||15.6 - 1000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant human GM-CSF. The factors listed below were prepared at 10ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.|
Factors assayed for cross-reactivity
|Recombinant human||G-CSF, M-CSF, FGF-4, FGF-5, FGF-6, IL-1α, IL-1β, IL-3, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, VEGF|
|Recombinant mouse||GM-CSF, IL-1α, IL-1β, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, TNF-α|
|Other proteins||bovine FGF acidic, bovine FGF basic, human PDGF, porcine PDGF, Human TGF-β1, porcine TGF-β1|
Data Analysis Assistance
We have partnered with MyAssays to offer you an easy to use and versatile tool to analyze the data you receive using our ELISA Kit. Click the link below to be directed to the data analysis tool provided by MyAssays specifically for BSKH1018.