Mouse ghcagons-like pepfide 1 (GLP-1) ELISA Kit instruction Kit name Mouse ghcagons-like pepfide 1 (GLP-1) ELISA Kit Intended use The kit is used to assay the content of Mouse ghcagons-like pepfide 1 (GLP-1) in Mouseserum, blood plasma and other related tissue liquid.Test principleThe kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) to assay the level of Mouse ghcagons-like pepfide 1 (GLP-1) in samples. Add Mouseghcagons-like pepfide 1 (GLP-1) to pre-coated Mouse ghcagons-like pepfide 1 (GLP-1) monoclonal antibody microelisa well, incubation; washing. Add HRP tagged Mouseghcagons-like pepfide 1 (GLP-1) antibodies. After another incubation and washing, removethe unbound enzyme, add Chromogen Solution A and B, the color of the liquid change intoblue, and the color finally become yellow at the effect of acid. The depth of the color ispositively correlated with concentration of the Mouse ghcagons-like pepfide 1 (GLP -1) insamples.Materials supplied1 Microelisa Stripplate 12well × 8strips 7 Chromogen Solution A 6mL2 Standard: 240pmol / L 0.6mL 8 Chromogen Solution B 6mL3 20 × wash solution 25mL 9 Stop Solution 6mL4 Standard diluent 6mL 10 Instruction 15 Sample diluent 6mL 11 Closure plate membrane 26 HRP-Conjugate Reagent 6mL 12 Sealed bags 1Note: Standard was diluent with Standard diluent followed by: 240, 120, 60, 30, 15, 7.5pmol / L. Materials required but not supplied 1. 37 ℃ incubator 2. Standard microplate reader 3. Precision pipettes and Disposable pipette tips 4. Distilled water5. Disposable tubes for sample dilution6. Absorbent paperAssay procedure1. Prepare: The kit takeing out from the environment of 2-8 ℃ should be balanced 30minutes at less in the room temperature before using.2. Diluent: Diluent the 20 × wash solution .3. Add standard and Sample: Set Standard wells, testing sample wells and blank wells. AddDiluted standard 50μl to standard well; Add Sample dilution 40μl to testing sample well which on Assay plate, then add testing sample 10μl (sample final dilute degree is 5times), blank well doesn't add anyting. 4. Incubation: Incubate 30 minutes at 37 ℃ in incubator. 5. Wash: Discard Liquid, drying, filling in diluted washing liquid to each well, oscillationfor 1 min, discard the washing liquid with absorbent paper Pat dry. Repeat three times, Pat dry. 6. Add HRP-conjugate reagent: Add HRP-conjugate reagent 50μl to each well, except theblank well. Mixing gently shaking, induced 30 minutes at 37 ℃. 7. Repeat step4.8. Repeat step59. Add chromogen solution A and B: Add chromogen solution A 50μl and chromogensolution B 50μl to each well. Gently mix, incubate for 15 min at 37 ℃. 10. Add Stop Solution: Add Stop Solution 50μl to each well, Stop the reaction (the blue colorchange to yellow immediately). 11. Take blank well as zero, measure the optical densit (OD) at 450 nm after adding StopSolution and within 15 min.12 . According to standard concentration and the corresponding OD values ​​calculatedstand ard curve linear regression equation, then the OD values ​​according to the sample onthe regression equation to calculate the corresponding sample concentration. It should be remembered that the sample has been diluted and its actual concentration should be multiplied by the total dilution.Specimen requirements1. Can't detect the samples which contain NaN3, because NaN3 inhibits HRP activity of thehorseradish peroxidase. 2. Extract as soon as possible after Specimen collection, Extracted according to the relevant literature, and should be experiment as soon as possible after the extraction. If it can notbe tested immediately, specimen can be kept in -20 ℃ to preserve, but repeated freezingand thawing should be avoided.3. The samples shoule be centrifugated dequately and no hemolysis or granule was allowed.Important notes1. The operation should be carried out in strict accordance with instructions and test resultsmust be based on microplate reader to determine readings shall prevail.2. If th e microelisa stripplate has not used up after open, it should be stored in the sealed bag 3. Recommended that all standard materials, test samples are doing double to minish the Experimental error. 4. Please multiply total dilution times when calculation. 5 times is the best dilute timeaccording to this ELISA Kit design. 5. If the testing material content in the sample is excessively high, please use Specialdilution to dilute certain multiple, then assay. 6. If the color too shallow, It may be appropriate to extend the substrate incubation time. 7. Add Sample with sampler each step and proofread its accuracy frequently to avoid the experimental error. In order to avoid cross-contamination, avoid to reusing the suctionhead and closure plate membrane. 8. Use the kit in validity, not mix the reagents of different batches. 9. Chromogen Solution B is light-sensitive, avoid prolonged exposure to light, Summary proceduresPreparing reagents, samples and standardAdd prepared sample and standard, incubate d 30 minutes at 37 ℃ Plate washed four times, adding HRP-Conjugate Reagent promote 30 minutes at 37 ℃ Plate washed four times, adding Chromogen Solution A and B promote 15 minutes at 37 ℃ Add stop solutionMeasure within 15minCalculationAssay range: 7.5-240pmol / LPackage size: 96 determinationsStorage: 2-8 ℃ .validity: six months. Shanghai Yuping Biological Technology Co., Ltd. mainly deals with various brand grade ELISA kits, with quality assurance and perfect after-sales service. And provide free generation testing. Serving universities and immunology research units. Technicians serve you better.
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