PDGF-AB ELISA Kit (Human) (OKBB00239)

Catalog No: OKBB00239

Size:96 Tests

Human PDGF-AB ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human PDGF-AB ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for PDGF-AB has been precoated onto 96-well plates. Standards(E.coli, (S87-T211)+(S82-T190)) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for PDGF-AB 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 absorbance of yellow is proportional to the human PDGF-AB amount of sample captured in plate.
• ELISA Kit Range: 31.2 pg/ml - 2,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	237	532	974	254	683	1205
  St. Dev.	11.6	27.7	62.4	13.97	45.8	95.2
  %CV	4.89	5.20	6.40	5.5	6.70	7.90

• ELISA Kit Component:
  • Lyophilized recombinant human MMP-7 standard: 10 ng/tube x 2
  • Anti-human MMP-7 Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human MMP-7 antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 31.2pg/ml-2000pg/ml
  Principle: Aviva’s human PDGF-AB ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human PDGF-AB specific-specific monoclonal antibodies were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells 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 human PDGF-AB amount of sample captured in plate.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
  experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
  marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
  using.
  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.
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum and plasma (heparin, EDTA)
• Sensitivity: <2 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• 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.
• Specificity: Natural and recombinant human PDGF-AB
• Immunogen: Expression system for standard: E.coli; Immunogen sequence: (S87-T211) + (S82-T190)
• Description: For quantitative detection of human PDGF-AB in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Target Info

• Gene Symbol: PDGFB
• Gene Full Name: platelet-derived growth factor beta polypeptide,platelet-derived growth factor alpha polypeptide
• Alias Symbols: PDGF A chain, PDGF B chain, PDGF1, PDGF2, PDGFA, PDGFB, Platelet derived growth factor AB heterodimer
• NCBI Gene Id: 5155; 5154
• Protein Name: Platelet-derived growth factor subunit B,Platelet-derived growth factor subunit A
• Description of Target: 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.
• Uniprot ID: P01127, P04085
• Protein Accession #: NP_002598.4
• Nucleotide Accession #: NM_002607.5