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Catalog No: OKBB00157
Size:96 Tests
Price: $466.00
SKU
OKBB00157
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Datasheets/ManualsClick here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Product Info
Predicted Species ReactivityHuman
ApplicationELISA
ELISA Kit Detection MethodColorimetric, OD450 nm
ELISA Kit Duration~ 3 Hours
ELISA Kit PrincipleAviva'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 Range125 pg/ml - 8,000 pg/ml
ELISA Kit Reproducibility
Intra-Assay PrecisionInter-Assay Precision
Sample123123
Mean (pg/mL)5213649562153235765618
St. Dev.39.6222.6269.840.96303.96348.3
%CV7.606.104.797.698.56.19
ELISA Kit Component
ComponentAmount
Lyophilized recombinant mouse Fractalkine standard10 ng/tube x 2
Anti-mouse Fractalkine Antibody Well Plate96 Wells
Sample diluent buffer30 mL
Biotinylated anti-mouse Fractalkine antibody130 uL, dilution 1:100
Antibody diluent buffer12 mL
Avidin-Biotin-Peroxidase Complex (ABC)130 uL, dilution 1:100
ABC diluent buffer12 mL
TMB color developing agent10 mL
TMB stop solution10 mL
Additional InformationRange: 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 StorageStore 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 SequenceNo detectable cross-reactivity with any other cytokine.
Reference1. 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.
SpecificityNatural and recombinant human HGF
ImmunogenExpression system for standard: NS0; Immunogen sequence: Q32-R494 (alpha) & V495-S728 (beta)
Gene SymbolHGF
Gene Full Namehepatocyte growth factor (hepapoietin A; scatter factor)
Alias SymbolsDFNB39, 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 Id3082
Protein NameHepatocyte growth factor
Description of TargetHepatocyte 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 IDP14210
Protein Accession #NP_000236.2
Nucleotide Accession #NM_000245.2
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