Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IGF-1 has been pre-coated onto a microplate. Standard, control, or sample and the working solution of Biotin-Conjugate are pipetted into the wells. Following incubation and wash steps, any IGF-1 present is bound by the immobilized antibody and the detection antibody specific for IGF-1 is binds to the combination of capture antibody-IGF-1 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 coloured product is formed in proportion to the amount of IGF-1 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 IGF-1 standard dilutions and IGF-1 sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Interleukin 6(IGF-1) is a multifunctional protein produced by lymphoid and non-lymphoid cells and by normal and transformed cells, including T cells, monocyte/macrophages, fibroblasts, hepatocytes, vascular endothelial cells, cardiac myxomas, bladder cell carcinomas, myelomas, astrogliomas and glioblastomas. The production of IGF-1 in these cells is regulated, either positively or negatively, by a variety of signals including mitogens, antigenic stimulation, lipopolysaccharides, IL-1, TNF, PDGF and viruses.
The human IGF-1 cDNA sequence predicts a protein of 212 amino acid (aa) residues in length with two potential N-glycosylation sites. The hydrophobic N-terminal 28 aa residue signal peptide is cleaved to produce a mature protein of 184 amino acids with four cysteine residues and a predicted molecular mass of 21 kDa. The mouse IGF-1 cDNA sequence shows a homology of 42% at the aa level when compared with the human sequence. On the basis of sequence similarity and gene structural motif similarity, IGF-1 can be grouped in a family of cytokines that also includes OSM, G-CSF, LIF, and CNTF. All of these cytokines are predicted to have a four-helix bundle structure similar to that found for growth hormone, suggesting that they all evolved from a common ancestral gene.
The effects of IGF-1 on different cells are numerous and varied. The effect on B cells is stimulation of differentiation and antibody secretion. IGF-1 also affects T cells, acting as a co-stimulant with sub-optimal concentrations of PHA or Con A to stimulate IL-2 production and IL-2 receptor expression.
IGF-1 exhibits growth factor activity for mature thymic or peripheral T-cells and reportedly enhances the differentiation of cytotoxic T-cells in the presence of IL-2 or IFN-γ. IGF-1 stimulates production of acute phase proteins by hepatocytes and has colony-stimulating activity on hematopoietic stem cells. IGF-1 has growth factor activities and will stimulate the growth of myeloma/hybridoma/ plasmacytoma cells, EBV-transformed B cells, keratinocytes and mesangial cells.
|SYNONYMS||IGF; MGF; IGFI; IGF-I|
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with antibody to human IGF-1 (812)||1 plate|
|Human IGF-1 standard lyophilized, 4000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-human IGF-1 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 30pg/mL.|
|ASSAY RANGE||62.50 - 4000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant human IGF-1. The factors listed below were prepared at 50ng/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||FGF-acidic, HGF , insulin, TGF-beta 1, TGF-beta 2, VEGF|
|Recombinant mouse||IGF-1, IGF-2|
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 BSKH1058.