Zebrafish bradykinin (BK) ELISA kit

Purpose of use:

This reagent kit is used for determinationzebrafishIn serum, plasma, and related liquid samplesbradykinin(BK)content.

Experimental Principle

This test kit uses the double antibody sandwich method for determinationspecimenChineseZebrafish bradykinin(BK)Level. Using purifiedZebrafish bradykinin(BK)Antibody coated microplates are used to prepare solid-phase antibodies, which are then sequentially added to the micropores coated with monoclonal antibodiesbradykinin(BK)，again withHRPmarkedbradykinin(BK)Antibody binding, forming antibodies-antigen-Enzyme linked antibody complexAfter thorough washingplussubstrateTMBColor rendering.TMBinHRPUnder the catalysis of enzymes, it is converted into blue, and under the action of acid, it is converted into the final yellow. The depth of color and the samplebradykinin(BK)Positive correlation. Using an enzyme-linked immunosorbent assay (ELISA) reader450nmMeasure absorbance at wavelength（ODValue),By standard curvecalculated sampleChineseZebrafish bradykinin(BK)Concentration.

Specimen requirements

1Extract the specimen as soon as possible after collection, according to relevant literature, and conduct experiments as soon as possible after extraction. If the experiment cannot be conducted immediatelyPlace the specimen in-20Stored at ℃, butshouldAvoid repeated freeze-thaw cycles

2. Cannot detect samples containing NaN3Due toNaN3Inhibition of horseradish peroxidase（HRP）Activity.

calculate

Using the concentration of the standard substance as the horizontal axis,ODThe value is on the vertical axis, and a standard curve is drawn on the coordinate paper based on the sampleODFind the corresponding concentration from the standard curve; Multiply againdilution factor; or using the concentration of standard substances andODCalculate the linear regression equation of the standard curve based on the value, and calculate the sample'sODSubstitute the value into the equation, calculate the sample concentration, and then multiply it bydilution factor, which is the actual concentration of the sample.

Precautions

1The reagent kit should be taken out of the refrigerated environment and equilibrated at room temperature15-30Minutes later, the enzyme label coated plate can be used. If it is not used up after opening, the Flat noodles should be stored in a sealed bag.

2．Concentrated detergent solutionpossiblythere will becrystallizationPrecipitation can be dissolved by heating in a water bath during dilutionWashing does not affect the results.

3Each step of sample addition should use a sampler and its accuracy should be regularly checked to avoid experimental errors. It is best to control the sampling time at once5Within minutes, if there are a large number of specimens, it is recommended to use a sampling gun to add samples.

4．Please make a standard curve at the same time as each measurement, preferably with a double hole. If the content of the substance to be tested in the specimen is too high (sample)ODValue greater than the first hole of the standard holeODPlease dilute the sample diluent to a certain multiple first（nMeasure again after doubling, please calculate lastmultiplied byTotal dilution factor（×n×5）.

5．The sealing film is only for one-time use to avoid cross contamination.

6Please store the substrate away from light.

7Strictly follow the instructions for operation, and the judgment of the test results must be based on the reading of the enzyme-linked immunosorbent assay reader.

8All samples, detergents, and various waste materials should be treated as infectious agents.

9Components from different batch numbers of this reagent must not be mixed.

Storage conditions and expiration date

1．Storage of reagent kit:；2-8℃.

2Validity period:6month