Instruction manual for rat (Rat) hepatocyte growth factor (HGF) ELISA test kit

This kit can only be used for scientific research, not for medical diagnostic rat (Rat) hepatocyte growth factor (HGF) ELISA test kit instructions for use. Principles of the test kit using double antibody one-step sandwich enzyme-linked immunosorbent assay ). To the coated microwells pre-coated with hepatocyte growth factor (HGF) antibody, add specimens,
Standards, HRP-labeled detection antibodies are incubated and washed thoroughly. The color is developed with the substrate TMB, which is converted into blue under the catalysis of peroxidase and into the final yellow under the action of acid. The color depth is positively correlated with the hepatocyte growth factor (HGF) in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the sample concentration was calculated.
Sample collection, processing and storage methods
1. Serum: Use a test tube that does not contain pyrogens and endotoxins. Avoid any cell irritation during the operation. After collecting blood, centrifuge at 3000 rpm for 10 minutes to quickly and carefully separate serum and red blood cells.
2. Plasma: EDTA, citrate or heparin anticoagulation. Take the supernatant by centrifugation at 3000 rpm for 30 minutes.
3. Cell supernatant: centrifuge at 3000 rpm for 10 minutes to remove particles and polymers.
4. Tissue homogenate: Add tissue to the right amount of saline and mash. Take the supernatant by centrifugation at 3000 rpm for 10 minutes.
5. Preservation: If the samples are not tested in time after collection, please aliquot according to the dosage and freeze in
-20 ℃, avoid repeated freezing and thawing, thaw at room temperature and ensure that the sample is thawed evenly and fully.
Bring your own items
1. Microplate reader (450nm)
2. High-precision sampler and tip: 0.5-10uL, 2-20uL, 20-200uL, 200-1000uL
3. Precautions for operation of 37 ℃ thermostat
1. Store the kit at 2-8 ° C and equilibrate at room temperature for 20 minutes before use. The concentrated washing liquid taken out of the refrigerator will have crystals, which is a normal phenomenon. The water bath is heated to completely dissolve the crystals before use.
2. The slats not used in the experiment should be immediately returned to the ziplock bag, sealed (dry at low temperature) and stored.
3. S0 standard with concentration of 0 can be regarded as negative control or blank; OD value after pretreatment.
Result judgment to draw a standard curve: In the Excel worksheet, the standard concentration is used as the abscissa, corresponding
The OD value is used as the ordinate, a linear regression curve of the standard product is drawn, and the concentration value of each sample is calculated according to the curve equation.
Kit performance
1. Accuracy: the correlation coefficient R between the linear regression of the standard product and the expected concentration is greater than or equal to
0.9900.
2. Sensitivity: The minimum detection concentration is less than 1.0 pg / mL.
3. Specificity: Does not cross-react with other soluble structural analogs.
4. Repeatability: The coefficients of variation within and between panels are less than 15%.
5. Storage: 2-8 ℃, protected from light and moisture.
6. Validity: 6 months disclaimer
1. The kit is for research use only, and should not be used for clinical or human experiments, otherwise all the consequences will be borne by the experimenter, and the company will not be responsible.
2. Strictly follow the instructions, and the experimenter violates the instructions, and the consequences will be borne by the experimenter.

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