HGF ELISA Kit (Human) (OKBB00157)

Catalog No: OKBB00157

Size:96 Tests

Human MET 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 HGF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for HGF has been precoated onto 96-well plates. Standards(NS0, Q32-R494( alpha) & V495-S728( beta)) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for HGF 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 HGF amount of sample captured in plate.
• ELISA Kit Range: 125 pg/ml - 8,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	521	3649	5621	532	3576	5618
  St. Dev.	39.6	222.6	269.8	40.96	303.96	348.3
  %CV	7.60	6.10	4.79	7.69	8.5	6.19

• ELISA Kit Component:
  • Lyophilized recombinant mouse Fractalkine standard: 10 ng/tube x 2
  • Anti-mouse Fractalkine Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-mouse Fractalkine 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 HGF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human HGF 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 HGF 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: Hepatocyte growth factor (HGF) is the most potent mitogen for mature parenchymal hepatocytes in primary culture, and seems to be a hepatotrophic factor that acts as a trigger for liver regeneration after partial hepatectomy and liver injury. HGF has a relative molecular mass (Mr) of 82,000 and is a heterodimer composed of a large alpha-subunit of Mr 69,000 and a small beta-subunit of Mr 34,000.1 The protein consists of 728 amino acid residues, including a possible signal peptide at the N-terminus.2 HGF may serve as a paracrine mediator to control placental development and growth.3 This growth factor may play an important role as a paracrine mediator of the proliferation of melanocytes and endothelial cells, as well as cells of epithelial origin.4 The gene encoding the human HGF is assigned to human chromosome 7.5 The standard product used in this kit is recombinant human HGF with the molecular mass of 60-70KDa.
• 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 (EDTA, citrate)
• Sensitivity: <5 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Nakamura, T.; Nishizawa, T.; Hagiya, M.; Seki, T.; Shimonishi, M.; Sugimura, A.; Tashiro, K.; Shimizu, S. Molecular
  cloning and expression of human hepatocyte growth factor. Nature 342: 440-443, 1989.
  2. Miyazawa, K.; Tsubouchi, H.; Naka, D.; Takahashi, K.; Okigaki, M.; Arakaki, N.; Nakayama, H.; Hirono, S.;
  Sakiyama, O.; Takahashi, K.; Gohda, E.; Daikuhara, Y.; Kitamura, N. Molecular cloning and sequence analysis of
  cDNA for human hepatocyte growth factor. Biochem. Biophys. Res. Commun. 163: 967-973, 1989.
  3. Kilby, M. D.; Afford, S.; Li, X. F.; Strain, A. J.; Ahmed, A.; Whittle, M. J. Localisation of hepatocyte growth factor and
  its receptor (c-met) protein and mRNA in human term placenta. Growth Factors 13: 133-139, 1996.
  4. Rubin, J. S.; Chan, A. M.-L.; Bottaro, D. P.; Burgess, W. H.; Taylor, W. G.; Cech, A. C.; Hirschfield, D. W.; Wong, J.;
  Miki, T.; Finch, P. W.; Aaronson, S. A. A broad-spectrum human lung fibroblast-derived mitogen is a variant of
  hepatocyte growth factor. Proc. Nat. Acad. Sci. 88: 415-419, 1991.
  5. Zarnegar, R.; Petersen, B.; DeFrances, M. C.; Michalopoulos, G. Localization of hepatocyte growth factor (HGF)
  gene on human chromosome 7. Genomics 12: 147-150, 1992.
• Specificity: Natural and recombinant human HGF
• Immunogen: Expression system for standard: NS0; Immunogen sequence: Q32-R494 (alpha) & V495-S728 (beta)
• Description: For quantitative detection of human HGF in cell culture supernatants, serum and plasma(EDTA, citrate).

Target Info

• Gene Symbol: HGF
• Gene Full Name: hepatocyte growth factor (hepapoietin A; scatter factor)
• Alias Symbols: DFNB39, F TCF, Fibroblast derived tumor cytotoxic factor, Hepapoietin A, Hepatocyte growth factor, Hepatocyte growth factor (hepapoietin A scatter factor, Hepatocyte growth factor beta chain, Hepatopoietin-A, Hgf, HGF_HUMAN, HGFB, HPTA, Lung fibroblast derived mitogen, Scatter factor, SF
• NCBI Gene Id: 3082
• Protein Name: Hepatocyte growth factor
• Description of Target: Hepatocyte growth factor (HGF) is the most potent mitogen for mature parenchymal hepatocytes in primary culture, and seems to be a hepatotrophic factor that acts as a trigger for liver regeneration after partial hepatectomy and liver injury. HGF has a relative molecular mass (Mr) of 82,000 and is a heterodimer composed of a large alpha-subunit of Mr 69,000 and a small beta-subunit of Mr 34,000.1 The protein consists of 728 amino acid residues, including a possible signal peptide at the N-terminus.2 HGF may serve as a paracrine mediator to control placental development and growth.3 This growth factor may play an important role as a paracrine mediator of the proliferation of melanocytes and endothelial cells, as well as cells of epithelial origin.4 The gene encoding the human HGF is assigned to human chromosome 7.5
• Uniprot ID: P14210
• Protein Accession #: NP_000236.2
• Nucleotide Accession #: NM_000245.2