Nanjing Senbega stock, quality assurance, price concessions, kits preferred Senbega. This reagent is for research use only: This kit is used to determine the content of nuclear factor-κB (NF-κB) in rat serum, plasma and related liquid samples. Experimental principle : The kit uses a double antibody sandwich assay to determine the level of nuclear factor-kappa B (NF-κB) in rats. The microplate was coated with purified rat nuclear factor-kappa B (NF-κB) antibody to prepare a solid phase antibody, and nuclear transcription factor-κB (NF-κB) was added to the microwell of the coated monoclonal antibody. The HRP-labeled nuclear transcription factor-κB (NF-κB) 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 the nuclear transcription factor-κB (NF-κB) in the sample. The absorbance (OD value) was measured by a microplate reader at a wavelength of 450 nm, and the content of rat nuclear transcription factor-κB (NF-κB) in the sample was calculated by a standard curve. Kit composition : Kit composition 48 hole configuration 96-well configuration save Instruction manual 1 copy 1 copy Sealing film 2 pieces (48) 2 pieces (96) sealed bag 1 1 Enzyme label coated plate 1×48 1×96 Store at 2-8 ° C Standard: 1350ng/L 0.5ml × 1 bottle 0.5ml × 1 bottle Store at 2-8 ° C Standard dilution 1.5ml × 1 bottle 1.5ml × 1 bottle Store at 2-8 ° C Enzyme standard reagent 3 ml × 1 bottle 6 ml × 1 bottle Store at 2-8 ° C Sample diluent 3 ml × 1 bottle 6 ml × 1 bottle Store at 2-8 ° C Developer A solution 3 ml × 1 bottle 6 ml × 1 bottle Store at 2-8 ° C Developer B solution 3 ml × 1 bottle 6 ml × 1 bottle Store at 2-8 ° C Stop solution 3ml × 1 bottle 6ml × 1 bottle Store at 2-8 ° C 20 times concentrated washing solution 20ml × 1 bottle 20ml × 1 bottle Store at 2-8 ° C Sample processing and requirements : 1. Serum: The blood is naturally coagulated for 10-20 minutes at room temperature and centrifuged for about 20 minutes (2000-3000 rpm). 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. Steps: Precautions: 10. In the case of an English manual, the English manual shall prevail. Calculation : The concentration of the standard is the abscissa and the OD is the ordinate. Draw a standard curve on the coordinate paper, depending on the OD of the sample The value is determined from the standard curve by the corresponding concentration; multiplied by the dilution Multiples; or use the concentration of the standard and the OD value to calculate the standard The linear regression equation of the quasi-curve, the OD value of the sample Substituting the equation, calculating the sample concentration, multiplying by dilution The multiple is the actual concentration of the sample. (This picture is for reference only) Kit performance: 1. The linear regression coefficient of the sample and the expected concentration correlation coefficient R value is 0.990 or more. 2. Within and within the batch should be less than 9% and 11% respectively examination range: 50 ng/L -1000 ng/L Storage conditions and expiration date: 1. Kit storage: 2-8 ° C. 2. Validity: 6 months
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