Brief description of the use of porcine adiponectin (ADP) ELSIA test kit - Database & Sql Blog Articles

Test - lowercase jpg

Kaixin micro test

Test probe P100-M3

EL-C1600N100013-B

Use of porcine adiponectin (ADP) ELSIA test kit:

This kit is used to determine the adiponectin (ADP) content in pig serum, plasma and related liquid samples.

Experimental principle of porcine adiponectin (ADP) ELSIA test kit:

This kit uses the double antibody sandwich method to determine the level of porcine adiponectin (ADP) in the specimen. Purified porcine adiponectin (ADP)

Antibody coated microwell

The plate is made into a solid phase antibody, and adiponectin (ADP) is sequentially added to the microwell of the coated monoclonal antibody, and then HRP is added.

Labeled adiponectin (ADP) resistance

Body-binding, forming an antibody-antigen-enzyme-labeled antibody complex, after thorough washing, adding a substrate

TMB color development. Catalysis of TMB in HRP enzyme

It is converted to blue and converted to the final yellow color by the action of an acid. The color depth and the adiponectin (ADP) in the sample are in the normal phase

turn off. Absorbance (OD) measured at 450 nm using a microplate reader

Value), the concentration of porcine adiponectin (ADP) in the sample was calculated from the standard curve.

Porcine adiponectin (ADP) ELSIA test kit

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 (240μg/L)

0.5ml × 1 bottle

3

Enzyme label coated plate

12 holes × 8

9

Standard dilution

1.5ml × 1 bottle

4

Sample diluent

6ml × 1 bottle

10

Instruction manual

1 copy

5

Developer A solution

6ml × 1 bottle

11

Sealing film

2 sheets

6

Developer B solution

6ml × 1 bottle

12

sealed bag

1

Pig adiponectin (ADP) ELSIA test kit specimen requirements

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.

2. Samples containing NaN3 could not be detected because NaN3 inhibited horseradish peroxidase (HRP) activity.

Porcine adiponectin (ADP) ELSIA test kit procedure

1. Dilution of standard: This kit provides one original standard, which can be diluted in a small tube according to the following chart.

120μg/L

Standard No. 5

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

60μg/L

Standard No. 4

Add 150 μl of standard dilution to 150 μl of standard #5

30μg/L

Standard No. 3

Add 150 μl of standard dilution to 150 μl of standard #4

15μg/L

Standard 2

Add 150 μl of standard dilution to 150 μl of Standard #3

7.5μg/L

Standard No. 1

150 μl of Standard 2 is added to 150 μl of standard dilution

2. Loading: Set blank holes separately (the blank control wells are not added with the sample and the enzyme standard reagent, the other steps are the same), the standard holes, and the sample holes 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: The plate was sealed with a sealing film and incubated at 37 ° C for 30 minutes.

4. Dosing: 30 times concentrated washing solution diluted with distilled water 30 times and used

5. Washing: Carefully remove the sealing film, discard the liquid, dry it, fill each well with the washing liquid, let stand for 30 seconds, then discard it, repeat 5 times, and pat dry.

6. Add enzyme: 50 μl of enzyme labeling reagent was added to each well, except for blank wells.

7. Incubation: The operation is the same as 3.

8. Washing: The operation is the same as 5.

9. Color development: add 50μl of color developer A, then add 50μl of color developer B, gently shake and mix, avoid light and develop color at 37 °C

15 minutes.

10. Termination: 50 μl of stop solution was added to each well to terminate the reaction (when 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.

Pig adiponectin (ADP) ELSIA test kit calculation

Taking the concentration of the standard as the abscissa and the OD as the ordinate, plotting the standard curve on the coordinate paper, according to the sample

The OD value is determined from the standard curve by the corresponding concentration; multiplied by the dilution factor; or the linear regression equation of the standard curve is calculated from the concentration of the standard substance and the OD value, and the OD value of the sample is substituted into the equation to calculate the sample concentration, and then multiplied The dilution factor is the actual concentration of the sample.

Porcine adiponectin (ADP) ELSIA test kit considerations

1. The kit should be taken out from the refrigerated environment and should be used at room temperature for 15-30 minutes before use. If the enzyme label is unsealed after opening, the slats 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 (sample OD value)

If it is larger than the OD value of the first hole of the standard product, please dilute it with a certain dilution (n times) with the sample diluent, and then multiply it by the total dilution factor (×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 instructions of the manual, the test results must be based on the microplate reader reading.

8. All samples, washings and various wastes should be treated as infectious materials.

9. The different batch components of this reagent must not be mixed.

10. If it is different from the English manual, the English manual shall prevail.

Storage conditions and expiration date of porcine adiponectin (ADP) ELSIA test kit

1. The kit is stored at: 2-8 °C.

2. Validity: 6 months