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|Application / Reactivity||Chicken||Cow (Bovine)||Dog (Canine)||Fish||Goat||Guinea Pig||Monkey||Mouse (Murine)||Pig (Porcine)||Rabbit||Rat (Rattus)||Sheep (Ovine)||Wild boar (Sus scrofa)|
|ELISA||1 ELISA Kits||9 ELISA Kits||13 ELISA Kits||1 ELISA Kits||1 ELISA Kits||2 ELISA Kits||2 ELISA Kits||20 ELISA Kits||6 ELISA Kits||10 ELISA Kits||19 ELISA Kits||1 ELISA Kits||1 ELISA Kits|
|Antigen||Hepatocyte Growth Factor (Hepapoietin A, Scatter Factor) (HGF) ELISA Kits|
Kits with alternative reactivity to:
|Method Type||Sandwich ELISA|
|Detection Range||31.2-2000 pg/mL|
|Minimum Detection Limit||31.2 pg/mL|
|7 references available|
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Product Details HGF ELISA KitTarget details Application Details Handling ProductDetails: References for HGF Kit (ABIN414383) Images
|Purpose||The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of HGF in Serum,Plasma,Tissue Homogenate,Cell Lysate,Cell Culture Supernatant,Biological Fluids|
|Sample Type||Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids|
This assay has high sensitivity and excellent specificity for detection of Hepatocyte Growth Factor (HGF).
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between Hepatocyte Growth Factor (HGF) and analogues was observed.|
|Material not included||
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|Alternative Name||HGF (HGF ELISA Kit Abstract)|
|Research Area||Cancer, Angiogenesis, Hormones, Growth Factors|
|Pathways||RTK Signaling, Carbohydrate Homeostasis, Glycosaminoglycan Metabolic Process, Synaptic Membrane, Signaling of Hepatocyte Growth Factor Receptor|
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Information on standard material:
|Sample Volume||100 μL|
|Assay Time||3 h|
|Protocol||The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Hepatocyte Growth Factor (HGF). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Hepatocyte Growth Factor (HGF). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Hepatocyte Growth Factor (HGF), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Hepatocyte Growth Factor (HGF) in the samples is then determined by comparing the O.D. of the samples to the standard curve.|
Serum: Allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000 × g. Assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Tissue Homogenates: The preparation of tissue homogenates will vary depending upon tissue type. For this assay, rinse tissues in ice-cold PBS (0.02mol/L,pH 7.0-7.2) to remove excess blood thoroughly and weigh before homogenization. Mince the tissues to small pieces and homogenize them in 5-10 mL of PBS with a glass homogenizer on ice (Micro Tissue Grinders work, too). Sonicate the resulting suspension with an ultrasonic cell disrupter or subject it to two freeze-thaw cycles to further break the cell membranes. Centrifugate the homogenates for 5 minutes at 5000 × g. Remove the supernate and assay immediately or aliquot and store at -20°C
Cell Lysate: Cells must be lysed before assaying according to the following directions. Adherent cells should be detached with trypsin and then collected by centrifugation (suspension cells can be collected by centrifugation directly). Wash cells three times in cold PBS. Resuspend cells in PBS (1×) and subject them to ultrasonication for 4 times (or Freeze cells at -20 °C. Thaw cells with gentle mixing. Repeat the freeze/thaw cycle for 3 times.) Centrifuge at 1500 × g for 10 minutes at 2 - 8°C to remove cellular debris.
Cell Culture Supernatant: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20 °C or -80 °C for later use. Avoid repeated freeze/thaw cycles.
Biological Fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20 °C or -80 °C for later use. Avoid repeated freeze/thaw cycles.
|Calculation of Results||
Average the duplicate readings for each standard, control, and samples and subtract the average zero standard optical density. Create a standard curve on log-log graph paper, with HGF concentration on the y-axis and absorbance on the x-axis. Draw the best fit straight line through the standard points and it can be determined by regression analysis. Using some plot software, for instance, curve expert 1.30, is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
In order to make the calculation easier, we plot the O.D. value of the standard (X-axis) against the known concentration of the standard (Y-axis), although concentration is the independent variable and O.D. value is the dependent variable. However, the O.D. values of the standard curve may vary according to the conditions of assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), plotting log of the data to establish standard curve for each test is recommended. Typical standard curve below is provided for reference only.
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hepatocyte Growth Factor (HGF) were tested 20 times on one plate, respectively.
|Restrictions||For Research Use only|
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|Precaution of Use||The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.|
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5 % within the expiration date under appropriate storage condition.
|Expiry Date||6 months|
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|Product cited in:||
Li, Chen, Wang, Lü, Huan, Fang, Han, Ge, Chen: "Changes in growth factor levels in the cerebrospinal fluid of autism patients after transplantation of human umbilical cord blood mononuclear cells and umbilical cord-derived mesenchymal stem cells." in: Genetics and molecular research : GMR, Vol. 15, Issue 2, 2016
Ahn, Chang, Sung, Yoo, Sung, Choi, Park: "Cell type-dependent variation in paracrine potency determines therapeutic efficacy against neonatal hyperoxic lung injury." in: Cytotherapy, Vol. 17, Issue 8, pp. 1025-35, 2015
Prystupa, Kiciński, Sak, Boguszewska-Czubara, Toruń-Jurkowska, Załuska: "Proinflammatory Cytokines (IL-1α, IL-6) and Hepatocyte Growth Factor in Patients with Alcoholic Liver Cirrhosis." in: Gastroenterology research and practice, Vol. 2015, pp. 532615, 2015
Russo: "Correlation Between Hepatocyte Growth Factor (HGF) and Gamma-Aminobutyric Acid (GABA) Plasma Levels in Autistic Children." in: Biomarker insights, Vol. 8, pp. 69-75, 2013
Russo, Pietsch: "Decreased Hepatocyte Growth Factor (HGF) and Gamma Aminobutyric Acid (GABA) in Individuals with Obsessive-Compulsive Disorder (OCD)." in: Biomarker insights, Vol. 8, pp. 107-14, 2013
Nagasawa, Kondou, Yasuda, Yamazaki, Nishida, Yamada: "[RAEB with ringed sideroblasts in a 3-month-old boy]." in: [Rinshō ketsueki] The Japanese journal of clinical hematology, Vol. 28, Issue 7, pp. 1181-6, 1988
Stare: "Marketing a nutritional "revolutionary breakthrough". Trading on names." in: The New England journal of medicine, Vol. 315, Issue 15, pp. 971-3, 1986
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