Enzyme Linked Immunoassay (ELISA) Kit for Trypanosoma marieii This kit is for research use only. Drug name: Generic name: Trypanosoma maritii ELISA kit Purpose of use: This kit is used for the qualitative determination of horse serum, plasma and related liquid samples of Trypanosoma cruzi. Experimental principle This kit uses an indirect method to determine Trypanosoma marieii in specimens. The microplate was coated with purified trypanosoma eosinii antibody to make a solid phase antibody. After incubating with the trypanosoma eosinophilus in the sample, biotin-labeled anti-IgG antibody was added and then combined with streptavidin-HRP , To form immune complexes, after thorough washing, add substrate TMB to develop color. TMB is converted into blue under the catalysis of HRP enzyme, and into the final yellow under the action of acid. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm and compared with the CUTOFF value to determine the presence or absence of Trypanosoma maritima in the specimen. The kit consists of 1 20 times concentrated washing solution 50ml × 1 bottle 8 sample diluent 6ml × 1 bottle 2 streptavidin-HRP 6ml × 1 bottle 9 negative control 0.5ml × 1 bottle 3 enzyme label coated plate 12 wells × 8 strips 10 positive control 0.5ml × 1 bottle 4 biotin-labeled anti-IgG antibody 6ml × 1 bottle 11 sealed bag 1 5 developer A solution 6ml × 1 bottle 12 3 sealing film 6 developer B 6ml × 1 / bottle 13 instructions 1 copy 7 Stopper 6ml × 1 vial sample requirements 1. The samples containing NaN3 cannot be detected because NaN3 inhibits horseradish peroxidase (HRP) activity. 2. Specimens are extracted as soon as possible after collection, and extraction is performed according to relevant literature, and experiments should be conducted as soon as possible after extraction. If the test cannot be carried out immediately, the specimen can be stored at -20 ℃, but repeated freezing and thawing steps should be avoided. 1. Numbering: Number the microwells corresponding to the samples in sequence, each plate should be set with 2 wells for negative control and 2 wells for positive control 1. Blank control (without sample, biotin-labeled anti-IgG antibody, streptavidin-HRP, the rest of the operation is the same) 1 well 2. Add sample: add negative control and positive control to the negative and positive control wells respectively 50μl. Then add 40μl of sample diluent to the sample well, and then add 10μl of the sample to be tested. Add the sample and add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, shake gently to mix, and incubate at 37 ℃ for 45 minutes. 3. Mixing solution: dilute 20-fold concentrated washing solution with distilled water 20-fold and reserve for use 4. Washing: carefully peel off the sealing film, discard the liquid, spin dry, fill each well with the washing solution, let it stand for 30 seconds and then discard , Repeat this 4 times, pat dry. 5. Add biotin-labeled anti-IgG antibody: Add 50 μl of biotin-labeled anti-IgG antibody to each well (except blank wells). Incubate at 37 ° C for 30 minutes. 6. Wash: operate the same as 4. 7. Add streptavidin-HRP: add 50 μl of streptavidin-HRP to each well (except the blank well), gently shake to mix, 37 Incubate at 30 ° C for 30 minutes. 8. Washing: The operation is the same as 4. 9. Color development: add 50μl of developer A to each well, then add 50μl of developer B, mix gently, and develop at 37 ° C for 15 minutes in the dark. 10. Termination: Add 50μl of stop solution to each well to stop the reaction (at this time, the blue color turns to yellow). 11. Determination: Measure the absorbance (OD value) of each well in sequence with the blank air conditioner at zero and 450 nm wavelength. The measurement should be carried out within 15 minutes after adding the stop solution. Judgment of results: Test validity: average value of positive control wells ≥1.00; average value of negative control ≤0.10 cut-off value (CUT OFF) calculation: cut-off value = average value of negative control wells +0.15 negative judgment: sample OD value <cut-off value (CUT OFF) for the negative positive judgment of Trypanosoma mayi: the sample OD value ≥ critical value (CUT OFF) is positive for Trypanosoma mayi 1. The operation is strictly in accordance with the instructions. The components of different batches of this reagent must not be mixed. 2. The kit should be equilibrated at room temperature for 15-30 minutes before being taken out of the refrigerated environment. If the enzyme label coated plate is unopened, the strip should be stored in a sealed bag. 3. Crystals may be precipitated in the concentrated washing liquid, which can be heated and dissolved in a water bath during dilution, and the results will not be affected during washing. 4. The sealing film is limited to one-time use to avoid cross-contamination. 5. Please keep the substrate away from light. 6. The result of the test must be determined by the reading of the microplate reader. When using dual wavelength detection, the reference wavelength is 630nm All samples, washing liquids and various wastes should be treated as infectious agents. The stop solution is 2M sulfuric acid, and you must pay attention to safety when using it. Specifications: 96 servings / box storage conditions and expiration date Kit storage :; 2-8 ℃. 2. Validity: 6 months Shanghai Yuping Biological Technology Co., Ltd. mainly deals with ELISA kits of various brands and grades, with quality assurance and perfect after-sales service. And provide free generation testing. 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