This kit can only be used for scientific research, not for medical diagnosis. 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.
Goat) Follicle stimulating hormone FSH) ELISA detection kit detection principle
The kit uses double antibody sandwich method enzyme-linked immunosorbent assay (ELISA). Coat the microtiter plate with purified antibody to make a solid-phase carrier. Add samples or standards and HRP-labeled detection antibody to the microwells coated with anti-follicle stimulating hormone (FSH) antibody in sequence. Color rendering. TMB 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 FSH in the sample. Measure the absorbance (OD value) at 450nm with a microplate reader to calculate the sample concentration.
Sample collection, processing and storage methods Sample collection, processing and storage methods Sample collection, processing and storage methods Sample collection, processing and storage methods
1. Serum: Use a test tube that does not contain pyrogens and endotoxins. Avoid any cell stimulation during the operation. After collecting blood, centrifuge at 3000 rpm for 10 minutes to separate the serum and red blood cells quickly and carefully.
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 Bring your own items Bring your own items Bring your own items
1. Microplate reader (450nm)
2. High-precision sampler and tip: 0.5-10uL, 2-20uL, 20-200uL, 200-1000uL
3. Cautions for 37 ℃ constant temperature oven operation Cautions for operation Cautions for operation Cautions for operation
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 standard is diluted before use. The diluted standard should be discarded immediately and cannot be stored. ng / mL.
3. The slats not used in the experiment should be immediately returned to the ziplock bag, sealed (low temperature drying) and stored. 2. Dilution of 20 × washing buffer: 1:20 dilution of distilled water.
4. Perform the incubation operation strictly in accordance with the time, amount of liquid and sequence indicated in the manual. Washing method Washing method Washing method Washing method
5.5.5.5. Shake all liquid components before use. 1. Manually wash the plate: throw away the liquid in the well, fill each well with the washing liquid, and after 1 minute, shake off the reagent kit to make up the reagent kit to make up the kit to make up the liquid in the well, pat it dry on the absorbent paper, so wash Plate 4-5 times.
Name name name name 96969696 hole configuration hole configuration hole configuration hole configuration 48484848 hole configuration hole configuration hole configuration hole configuration Remarks Remarks Remarks 2. Automatic plate washing machine: Inject 350μL of washing solution into each well, soak for 1min, wash the plate 4-5 times.
Microwell microplate 12-well × 8 strips 12-well × 4 strips No operation steps Operation steps Operation steps Operation steps Standard (120ng / mL) 0.5mL 0.5mL Dilute according to the instructions Sample dilution 6mL 3mL None
1. Remove the required slats from the aluminum foil bag after 20 minutes of room temperature equilibration, and the remaining slats are sealed and placed with the standard diluent 6mL 3mL
Detection antibody-HRP 6mL 3mL without returning to 4 ℃.
20 × Wash buffer 25mL 15mL Dilute according to the instructions Substrate A 6mL 3mL None
2. In addition to the blank wells, add 50 μL of standard products of different concentrations to each standard well; sample substrate B to be tested 6 mL 3 mL without stop solution 6 mL 3 mL without wells First add 10 μL of the sample to be tested, and then add 40 μL of the sample diluent 5
Sealing film 2 sheets 2 sheets without instructions 1 copy 1 copy without magnification); then add a horseradish peroxidase (HRP) standard self-sealing bag 1 standard 1 and a memorized detection antibody 50μL to each well of the standard and sample wells Seal the reaction well with a sealing plate film, 37 ° C water bath or thermostat temperature reagent preparation reagent preparation reagent preparation reagent preparation reagent preparation incubation for 50min.
1. Standard product: Standard product (120ng / mL) is multiplied with standard product diluent before the experiment
3. Discard the liquid, pat dry on absorbent paper, fill each well with washing liquid, let stand for 1min, shake off the dilution, and get 6 concentration points (including standard points): 120, 60, 30, 15, 7.5, 3.75
Wash the liquid and pat dry on the absorbent paper, and repeat washing the plate 5 times in this way (you can also use the washing machine according to the instructions 4. Repeatability: The coefficient of variation within and between the plates is less than 15%.
Book operation wash board). 5. Detection range: 0-120ng / mL
4. Add 50μL of substrate A and B to each well, mix gently by shaking, and incubate at 37 ° C in the dark for 15 minutes. 6. Storage: 2-8 ℃, protected from light and moisture.
5. Add 50μL of stop solution to each well, and measure the well's 7. at the wavelength of 450nm within 15min. Validity period: 6 months
OD value. Disclaimer Disclaimer Disclaimer Disclaimer Disclaimer Result Judgment Result Judgment Result Judgment Result Judgment 1. The kit is for research use only and should not be used in clinical or human experiments, otherwise
1. The OD value of each standard and sample should be subtracted from the OD value of the blank well. The experimenter shall bear all the consequences, and our company shall not be responsible.
2. Draw the standard curve: In the Excel worksheet, use the concentration of the standard product as the abscissa. For the operation strictly in accordance with the instructions, the experimenter violates the instructions, and the result should be the ordinate of the OD value, and the linear regression of the standard product is drawn. Curve, calculate each commitment according to the curve equation.
The sample concentration value, multiplied by the dilution factor, is the sample concentration.
Kit performance Kit performance Kit performance 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 No. 6 standard.
3. Specificity: Does not cross-react with other soluble structural analogs.
Beijing Qisong Biological Technology Co., Ltd. is a professional agent for many well-known brands in the international life science field. Committed to the sales and promotion of various ELISA kits, immunohistochemical kits, primary and secondary antibodies, cytokines, biological reagents, pipettes, and consumables. Company website: Order phone
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