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EK000035-EK0484: Human PDGF-AB ELISA Kit
Range 31.2pg/ml-2000pg/ml
Sensitivity < 2 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of human PDGF-AB in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
The platelet-derived growth factor (PDGF) is a mitogen derived from human platelets consisting of two related polypeptides termed A and B chains.1 The genes for PDGF A chain, B chain/c-sis, and the PDGF receptor are expressed in human malignant glioma cell lines.2 Normal human endothelial cells in culture express the B chain of PDGF, and that endothelial-derived PDGF B chain is synthesized as a predicted precursor polypeptide of Mr 27,281.3 The entire B chain of PDGF is highly (96%) homologous to a portion of p28sis, the transforming protein of simian sarcoma virus (SSV). It has been suggested that p28sis exerts its transforming potential by mimicking the growth promoting activity of PDGF and stimulating the cell in an autocrine manner.1 PDGF A-chain precursor polypeptide is assigned to the proximal long arm of chromosome 7, band q11.23.4 The human homolog (PDGF B-chain/c-sis) of the transforming gene of simian sarcoma virus is assigned to chromosome 22.5 The standard product used in this kit is recombinant human PDGF-AB with the molecular mass of 27KDa.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Kelly, J. D.; Raines, E. W.; Ross, R.; Murray, M. J. The B chain of PDGF alone is sufficient for mitogenesis. EMBO J. 4: 3399-3405, 1985.
2. Hermansson, M.; Nister, M.; Betsholtz, C.; Heldin, C.-H.; Westermark, B.; Funa, K. Endothelial cell hyperplasia in human glioblastoma: coexpression of mRNA for platelet-derived growth factor (PDGF) B chain and PDGF receptor suggests autocrine growth stimulation. Proc. Nat. Acad. Sci. 85: 7748-7752, 1988.
3. Collins, T.; Ginsburg, D.; Boss, J. M.; Orkin, S. H.; Pober, J. S. Cultured human endothelial cells express platelet-derived growth factor B chain: cDNA cloning and structural analysis. Nature 316: 748-750, 1985.
4. Stenman, G.; Rorsman, F.; Betsholtz, C. Sublocalization of the human PDGF A-chain gene to chromosome 7, band q11.23, by in situ hybridization. Exp. Cell Res. 178: 180-184, 1988.
5. Dalla-Favera, R.; Gallo, R. C.; Giallongo, A.; Croce, C. Chromosomal localization of the human homolog (c-sis) of the simian sarcoma virus onc gene. Science 218: 686-688, 1982.
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