Chicken albumin (ALB) ELISA test kit instructions - Database & Sql Blog Articles

Kit instruction manual

This kit is for research use only. examination range:

96T

1.0ng/ml

-

20ng/ml

Chicken Albumin (ALB) ELISA Test Kit Instructions Purpose: This kit is used to determine albumin (ALB) content in chicken serum, plasma and related liquid samples. Experimental principle This kit uses the double antibody sandwich method to determine the level of chicken albumin (ALB) in the specimen. The microplate was coated with purified chicken albumin (ALB) antibody to prepare a solid phase antibody, and albumin (ALB) was sequentially added to the microcapsules of the coated monoclonal antibody, and then combined with HRP-labeled albumin (ALB) antibody. The antibody-antigen-enzyme-labeled antibody complex is formed, and after thorough washing, the substrate TMB is added 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 albumin (ALB) in the sample. The absorbance (OD value) was measured at 450 nm using a microplate reader, and the chicken albumin (ALB) concentration in the sample was calculated from a standard curve. Chicken albumin (ALB) ELISA test kit instruction kit composition 1 30 times concentrated washing solution 20ml × 1 bottle 7 stop solution 6ml × 1 bottle 2 enzyme standard reagent 6ml × 1 bottle 8 standard product (40

Ng/ml

) 0.5ml × 1 bottle 3 Enzyme label coating plate 12 holes × 8 strips 9 standard dilutions 1.5ml × 1 bottles 4 sample dilutions 6ml × 1 bottles 10 instructions 1 part 5 color reagent A liquid 6ml × 1 bottle 11 Sealing film 2 sheets 6 developer B liquid 6ml × 1 bottle 12 sealed bag 1 specimen requirement 1. 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. Cannot detect

NaN3 sample, due to

NaN3 inhibits horseradish peroxidase (HRP) activity. Procedure 1. Dilution of standard: This kit provides one original standard, which can be diluted in a small tube according to the following chart. 20

Ng/ml

Add 150μl standard dilution 10 to the original standard of 150μl standard 5

Ng/ml

Add No. 4 standard 150μl of No. 5 standard to 150μl standard dilution 5.0

Ng/ml

Add No. 4 standard 150μl of No. 4 standard to 150μl standard dilution 2.5

Ng/ml

Add No. 2 standard 150μl of No. 3 standard to 150μl standard dilution 1.25

Ng/ml

Add No. 1 standard 150μl No. 2 standard to 150μl standard dilution 2. Add sample: blank wells (blank control well without sample and enzyme standard reagent, the other steps are the same), standard well, chicken albumin (ALB) ELISA Test Kit Instructions Sample wells to be tested. Accurately load 50 μl of the standard on the enzyme-labeled plate, add 40 μl of the sample dilution to the well to be tested, and then add 10 μl of the sample to be tested (the final dilution of the sample is 5 times). Add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, and shake gently to mix. 3. Incubation: After sealing with a sealing film, incubate at 37 °C

3

0 minutes. 4. Liquor: Dilute 30 times concentrated washing solution with distilled water 30 times and use it. 5. Wash: carefully remove the sealing film, discard the liquid, dry it, fill each well with washing liquid, let stand for 30 seconds and discard it. , repeat this 5 times, pat dry. 6. Add enzyme: Add 50 μl of enzyme labeling reagent to each well, except for blank wells. 7. Incubation: operation is the same as 3. 8. Washing: operation is the same as 5. 9. Color development: add 50μl of coloring agent to each well, then add coloring agent B50μl, gently shake and mix, avoid light and develop color at 37°C 10 minutes

.

10. Termination: 50 μl of stop solution was added to each well to stop the reaction (the blue color turned yellow). 11. Measurement: The absorbance (OD value) of each well was measured sequentially with a blank air conditioner of zero and a wavelength of 450 nm. The measurement should be carried out within 15 minutes after the addition of the stop solution. Summary of operation procedure: Calculate the concentration of the standard as the abscissa and the OD value as the ordinate. Draw a standard curve on the coordinate paper, and find the corresponding concentration from the standard curve according to the OD value of the sample; multiply by the dilution factor; or Calculate the linear regression equation of the standard curve by using the concentration of the standard and the OD value. Substituting the OD value of the sample into the equation, calculating the sample concentration, and multiplying by the dilution factor, is the actual concentration of the sample. Chicken albumin (ALB) ELISA test kit instructions 1. The kit should be taken out from the refrigerated environment and allowed to equilibrate for 15-30 minutes at room temperature. If the enzyme label is unsealed after unsealing, the strip should be stored in a sealed bag. 2. Concentrated washing liquid may crystallize out. When diluted, it can be heated and dissolved in a water bath. The washing will not affect the result. 3. The sampler should be used for each step, and the accuracy should be corrected frequently to avoid test errors. It is best to control the loading time within 5 minutes. If the number of specimens is large, it is recommended to use a gun. 4. Please make a standard curve at the same time of each measurement, it is best to make a double hole. If the content of the substance to be tested in the specimen is too high (the OD value of the sample is larger than the OD value of the first hole of the standard well), please first dilute the sample dilution with a certain multiple (n times) and then measure it. When calculating, multiply the total dilution by the total dilution. Multiple (×

n × 5).

5. The sealing film is intended for single use only to avoid cross-contamination. 6. Please keep the substrate away from light. 7. Strictly follow the operation of the manual, the test results must be based on the reading of the microplate reader. All samples, washings and various wastes should be treated as infectious materials. 9. The different batch components of this reagent must not be mixed. Storage conditions and validity period 1. The kit is stored at: 2-8 °C. 2. Validity: 6 months

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