Human glycocholic acid (CG) enzyme-linked immunosorbent assay (ELISA) kit instruction manual

This reagent is for research use only: This kit is used to determine the content of glycocholic acid (CG) in human serum, plasma and related liquid samples. Experimental principle: This kit uses the double antibody sandwich method to determine the level of human glycocholic acid (CG) in the specimen. The microplate was coated with purified human glycocholic acid (CG) antibody to prepare a solid phase antibody, and glycocholic acid (CG) was added to the microcapsule of the coated monoclonal antibody, followed by HRP-labeled glycocholic acid (CG). The antibody binds to form an antibody-antigen-enzyme-labeled antibody complex, which is thoroughly washed and then added to the substrate TMB for color development. TMB is converted to blue under the catalysis of HRP enzyme and converted to the final yellow color by the action of an acid. The color depth is positively correlated with glycocholic acid (CG) in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the content of human glycocholic acid (CG) in the sample was calculated from a standard curve. Kit composition: Kit composition 48-well configuration 96-well configuration Storage instructions 1 part 1 part sealing film 2 pieces (48) 2 pieces (96) Sealed bag 1 1 enzyme label coated plate 1 × 48 1 × 96 2 -8 ° C preservation standard: 1350 ng / ml 0.5ml × 1 bottle 0.5ml × 1 bottle 2-8 ° C preservation standard dilution 1.5ml × 1 bottle 1.5ml × 1 bottle 2-8 ° C preservation enzyme standard reagent 3 ml × 1 bottle 6 ml × 1 bottle 2-8 ° C Preservation sample dilution 3 ml × 1 bottle 6 ml × 1 bottle 2-8 ° C Preservation developer A solution 3 ml × 1 bottle 6 ml × 1 bottle 2-8 ° C preservation Reagent B liquid 3 ml × 1 bottle 6 ml × 1 bottle 2-8 ° C Preservation solution 3 ml × 1 bottle 6 ml × 1 bottle 2-8 ° C Preservation concentrated washing solution (20ml × 20 times) × 1 bottle (20ml × 30 times) × 1 bottle 2-8 ° C preservation sample processing and requirements: 1. Serum: room temperature blood solidification for 10-20 minutes, centrifugation for about 20 minutes (2000-3000 rev / min). The supernatant is carefully collected, and if precipitation occurs during storage, it should be centrifuged again. 2. Plasma: EDTA or sodium citrate should be selected as an anticoagulant according to the requirements of the specimen. After mixing for 10-20 minutes, centrifuge for about 20 minutes (2000-3000 rpm). The supernatant is carefully collected, and if a precipitate forms during storage, it should be centrifuged again. 3. Urine: Collect with a sterile tube and centrifuge for about 20 minutes (2000-3000 rpm). The supernatant is carefully collected, and if a precipitate forms during storage, it should be centrifuged again. The chest and ascites and cerebrospinal fluid are referred to. 4. Cell culture supernatant: When detecting secreted components, collect them in a sterile tube. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. When the components in the cells were detected, the cell suspension was diluted with PBS (pH 7.2-7.4), and the cell concentration reached about 1 million/ml. By repeated freezing and thawing, the cells are destroyed and the intracellular components are released. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again. 5. Tissue specimen: After cutting the specimen, weigh the weight. Add a certain amount of PBS, pH 7.4. It was quickly frozen and stored in liquid nitrogen for use. The specimen still maintains a temperature of 2-8 ° C after melting. A certain amount of PBS (pH 7.4) was added, and the specimen was homogenized by hand or homogenizer. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. One part of the package is to be tested, and the rest is frozen for use. 6. The specimens should be extracted as soon as possible after collection, and the extraction should be carried out according to the 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 °C, but repeated freezing and thawing should be avoided. 7. Samples containing NaN3 cannot be detected because NaN3 inhibits horseradish peroxidase (HRP) activity.

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