Shanghai Hengdailao Trading Co., Ltd. is mainly engaged in the research and development and sales of laboratory equipment, immunology, molecular biology and conventional biochemical reagents. And agent sales of many well-known foreign brands, is committed to providing first-class scientific research reagents and perfect technical services for the vast number of universities, research institutes and enterprises. This kit can only be used for scientific research, not for medical diagnosis. Rabbit thymosin β4 (Tβ4) quantitative detection kit (ELISA) user's Guide ã€Rabbit Thymosin β4 (Tβ4) Elisa Kit Kit Name】 Rabbit thymosin β4 (Tβ4) quantitative detection kit (ELISA) ã€Rabbit Thymosin β4 (Tβ4) Elisa Kit Kit Usage】 Quantitative detection of thymosin β4 (Tβ4) in rabbit serum, plasma, tissues and related fluid samples. [Rabbit Thymosin β4 (Tβ4) Elisa kit detection principle] This kit uses double-antibody two-step sandwich enzyme-linked immunosorbent assay (ELISA). The standard and the sample to be tested are added to the transparent enzyme label coated plate of rabbit thymosin β4 (Tβ4) monoclonal antibody pre-coated. After incubation for a sufficient time, the unbound components are washed to remove, and then the enzyme label working solution is added. After incubation for a sufficient time, washing 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 becomes yellow under the action of acid. The concentration of (Tβ4) was positively correlated. The OD value was measured at a wavelength of 450 nm. Based on the OD value of the standard and the sample, the content of rabbit thymosin β4 (Tβ4) in the sample was calculated. [Rabbit Thymosin β4 (Tβ4) Elisa Kit Kit Composition] 1 Enzyme coated plate 12 holes × 8 7 Developer A liquid 6mL 2 Standard 0.3mL × 6 tubes 8 Developer B liquid 6mL 3 20 times concentrated washing liquid 25mL 9 Stop solution 6mL 4 Sample diluent 6mL 10 Instructions 1 serving 5 Special diluent 6mL 11 Sealing film 2 sheets 6 Enzyme reagent 6mL 12 sealed bag 1 Remarks: The concentration of standard products (No. 1 → No. 6) is: 160, 80, 40, 20, 10, 5 mg / L. [Rabbit Thymosin β4 (Tβ4) Elisa kit required but not provided reagents and equipment] 1. 37 ℃ thermostat 2. Standard specification microplate reader 3. Precision pipettes and disposable tips 4. Distilled water 5. Disposable test tubes 6. Absorbent paper ã€Rabbits Thymosin β4 (Tβ4) Elisa Kit 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 standard product; first add 10μL of the sample to be tested, then add 40μL of sample diluent (that is, the sample is diluted 5 times); the blank control well is not added. 4. Incubation: Incubate in a 37 ° C water bath or thermostat for 30 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 plate 4 times (you can also use the washing machine to press Instructions for washing the board). 6. Add enzyme-labeled working solution: add 50μL of enzyme-labeled working solution to each well, without adding blank control wells. 7. Incubation: Incubate in a 37 ° C water bath or thermostat for 30 minutes. 8. Plate washing: Discard the liquid, pat dry on the absorbent paper, fill each hole with the washing liquid, let stand for 1min, shake off the washing liquid, pat dry on the absorbent paper, repeat the plate washing 4 times (you can also use the washing machine to press Instructions for washing the board). 9. Color development: add 50μL of developer A solution to each well, and then add 50μL of developer B solution. Mix with a plate mixer for 30s (or shake gently by hand for 30s), and avoid color development at 37 ℃ for 15min . 10. 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). 11. 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. 12. 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. [Rabbit thymosin β4 (Tβ4) Elisa kit sample requirements] 1. The sample cannot 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. ã€Precautions for rabbit thymosin β4 (Tβ4) Elisa kit】 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 5-160mg / L. If it exceeds this range, it is calculated from the extension of the standard curve. It is not used as an accurate quantitative result. ), 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. [Rabbit thymosin β4 (Tβ4) Elisa kit operating procedures summary] Prepare reagents, samples and standards Add prepared samples and standards, and react at 37 ℃ for 30 minutes Wash the plate 4 times, add the enzyme reagent, and react at 37 ℃ for 30 minutes Wash the plate 4 times, add color developing solutions A and B, and develop at 37 ℃ for 15 minutes Add stop solution Read OD value within 15 minutes Calculation [Rabbit Thymosin β4 (Tβ4) Elisa kit detection range] 5mg / L – 160mg / L ã€Rabbit Thymosin β4 (Tβ4) Elisa Kit Specifications】 96T / box ã€Storage of Rabbit Thymosin β4 (Tβ4) Elisa Kit】 Store at 2-8 ℃, protected from light and moisture. [Rabbit thymosin β4 (Tβ4) Elisa kit expiration date] 6 months
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Rabbit Thymosin β4 (Tβ4) Quantitative Detection Kit (ELISA) Instructions for Use