Mouse Follicle Stimulating Hormone (FSH) Quantitative Detection Kit (ELISA) Instruction Manual [Kit Name] Mouse Follicle Stimulating Hormone (FSH) Quantitative Detection Kit (ELISA) [Kit Use] To quantitatively detect mouse serum, The content of follicle stimulating hormone (FSH) in plasma and related fluid samples. [Detection principle] This kit adopts double antibody two-step sandwich enzyme-linked immunosorbent assay (ELISA). Add the standard and the test sample to the transparent enzyme label coated plate pre-coated with mouse follicle stimulating hormone (FSH) antibody. After incubation for a sufficient time, wash to remove unbound components, and then add the enzyme label working solution. After sufficient incubation time, wash to remove unbound components. Substrates A and B are added in sequence, and the substrate (TMB) is converted into a blue product catalyzed by horseradish peroxidase (HRP), which turns yellow under the action of acid. The concentration of FSH is positively correlated. The OD value is measured at a wavelength of 450 nm. Based on the OD value of the standard and the sample, the content of mouse follicle stimulating hormone (FSH) in the sample is calculated. [Composition of the kit] 1 enzyme label coating plate 12 wells × 8 strips 7 Developer A solution 6mL 2 Standard product: 24mIU / mL 0.6mL 8 Developer B solution 6mL 3 20-fold concentrated washing solution 25mL 9 Stop solution 6mL 4 standard product Diluent 6mL 10 Instructions 1 copy 5 Sample diluent 6mL 11 Sealing film 2 sheets 6 Enzyme label reagent 6mL 12 Sealed bag 1 Remarks: Standards are diluted with standard dilutions in order as follows: 24, 12, 6, 3, 1.5 , 0.75mIU / mL [reagents and equipment not needed] 1, 37 ℃ incubator 2, standard specification microplate reader 3, precision pipette and disposable pipette tip 4, distilled water 5, disposable test tube 6, water absorption Paper [Operation Steps] 1. Preparation: Remove the reagent kit from the refrigerator and re-equilibrate at room temperature for 30 minutes. 2. Mixing solution: dilute the 20-fold concentrated washing solution with distilled water to the original one. 3. Add standard products and samples to be tested: take a sufficient number of enzyme-coated plates and fix them to the frame. Set up standard wells, sample wells to be tested and blank control wells, record the positions of each well in the standard wells. Add 50μL of the standard; add 10μL of the sample to be tested to the sample well, then add 40μL of the sample diluent (that is, the sample is diluted 5 times); add 50μL of the enzyme-labeled working solution to both the standard and the sample well; . 4. Incubation: Incubate in a 37 ° C water bath or thermostat for 60 minutes. 5. Wash the plate: discard the liquid, pat dry on the absorbent paper, fill each well with the washing liquid, let stand for 1min, shake off the washing liquid, pat dry on the absorbent paper, repeat the washing of the plate 5 times (you can also use the washing machine to press Instructions for washing the board). 6. Color development: add 50 μL of developer A solution to each well, then add 50 μL of developer B solution, and develop color at 37 ° C in the dark for 15 min. 7. Termination: Remove the enzyme labeling plate and add 50μL of stop solution to each well to stop the reaction (the color changes from blue to yellow). 8. Determination: Zero the blank holes, and within 15 minutes after termination, measure the absorbance (OD value) of each well with a wavelength of 450 nm. 9. Calculation: According to the concentration of the standard product and the corresponding OD value, calculate the linear regression equation of the standard curve, and then calculate the corresponding sample concentration on the regression equation according to the OD value of the sample. You can also use various application software to Calculation. The final concentration is the actual measured concentration times the dilution factor. [Sample requirements] 1. The sample must not contain sodium azide (NaN3), because sodium azide (NaN3) is an inhibitor of horseradish peroxidase (HRP). 2. The specimen should be extracted as soon as possible after collection. The extraction should be carried out according to relevant literature. The experiment should be carried out as soon as possible after extraction. If the test cannot be performed immediately, the specimen can be stored at -20 ℃, but repeated freezing and thawing should be avoided. 3. The sample should be fully centrifuged, without hemolysis and particles. [Notes] 1. The experiment is carried out in strict accordance with the instructions, and the result of the experiment must be determined by the reading of the microplate reader. 2. If the enzyme-labeled coated board is not used up after opening, it should be put into a sealed bag and added with desiccant immediately. 3. It is recommended that all standards, samples and blank controls be tested in duplicate, and the average value is taken to reduce the experimental error. 4. Keep in mind that the sample has been diluted 5 times. The calculation result is multiplied by 5 to obtain the actual concentration of the sample. 5. The quantitative range of this kit is 0.75-24mIU / mL. If it exceeds this range, it is calculated from the extension of the standard curve. It is not used as an accurate quantitative result. Inside), multiplied by the total dilution factor is the final concentration of the sample. 6. If the color is too light, the substrate incubation time can be extended properly. 7. In order to avoid cross-contamination, the tips, samples and blank controls should be replaced with a new one for each addition; the common components such as enzyme working solution, sample diluent and substrate should be cantilevered, and they should not touch the microwells. ; Do not reuse the sealing film. 8. The kits are used within the warranty period, and different batches of reagents should not be mixed. 9. Substrate B is sensitive to light and avoid prolonged exposure to light. [Summary of operating procedures] Prepare reagents, samples and standards Add the prepared samples and standards, wash the plate 4 times at 37 ° C for 30 minutes, add the enzyme reagent, wash the plate 4 times at 37 ° C for 30 minutes and add the coloring solution A, B, color development at 37 ° C for 15 minutes. Add stop solution and read OD value within 15 minutes. [Detection range] 0.75-24mIU / mL [Specification] 96 servings / box [Storage] 2-8 ° C, protected from light and moisture . [Validity period] 6 months
Eight Function Bed,Adjustable Hospital Bed,Full Electric Hospital Bed,Patient Hospital Bed
Hebei Boxin Rehabilitation Equipment Co., Ltd , https://www.cnsmartcare.com