Human β - glucan binding protein (β - GBP) ELISA kit

Human β - glucan binding protein (β - GBP) ELISA kit

Specimen requirements
1. Sample processing: (1) After collecting the water sample, it is repeatedly frozen and thawed three times at -20 ℃, and then filtered through glass fiber for future reference
(2) Extract the tissue sample with butanol: methanol: water (5:25:70 V: V: V), or by phase extraction
Literature extraction should be carried out, and experiments should be conducted as soon as possible after extraction. If the experiment cannot be conducted immediately,
The specimen can be stored at -20 ℃ for future reference
2. Samples containing NaN3 cannot be detected because NaN3 inhibits the activity of horseradish peroxidase (HRP).

Human β - glucan binding protein (β - GBP) ELISA kitoperating steps
1. Sample addition: Set up standard wells and blank wells respectively (blank control wells do not add samples or enzyme-linked immunosorbent assay reagents, the rest of the steps are the same)
Sample hole to be tested. Add 50 microliters to the standard well on the enzyme-linked immunosorbent assay (ELISA) coated plate, and first add 40 μ l of sample diluent to the test sample well,
Then add 10 μ l of the sample to be tested (with a final dilution of 5 times). Add the sample to the bottom of the enzyme-linked immunosorbent assay (ELISA) plate well
Measure without touching the wall of the hole, gently shake and mix well.
2. Enzyme addition: Add 50 μ l of enzyme labeled reagent to each well, except for blank wells.
3. Incubation: Cover the plate with a sealing film and incubate at 37 ℃ for 60 minutes.
4. Solution preparation: Dilute 30 times the concentrated washing solution with distilled water and set aside for later use
5. Washing: Carefully remove the sealing film, discard the liquid, shake dry, fill each hole with washing solution, let it stand for 30 seconds, and then discard it
Repeat 5 times and pat dry.
6. Color development: Add 50 μ l of color developing agent A50 to each well, then add 50 μ l of color developing agent B50, gently shake and mix well, and develop color at 37 ℃ in the dark
15 minutes.
2
7. Termination: Add 50 μ l of termination solution to each well to terminate the reaction (at this point, blue will turn yellow).
8. Measurement: Zero the blank well and measure the absorbance (OD value) of each well in sequence at a wavelength of 450nm. The determination should be terminated upon addition
Perform within 15 minutes after the liquid is dissolved.

calculate
Draw a standard curve on a coordinate paper with the concentration of the standard substance as the horizontal axis and the OD value as the vertical axis. Based on the sample's
The OD value is determined by the standard curve to determine the corresponding concentration; Multiply by the dilution factor again; Or calculate the standard using the concentration and OD value of the standard substance
The linear regression equation of the quasi curve involves substituting the OD value of the sample into the equation to calculate the sample concentration, which is then multiplied by the dilution factor,
It is the actual concentration of the sample.

Precautions
1. The reagent kit should be taken out of the refrigerated environment and equilibrated at room temperature for 15-30 minutes before use. If the enzyme-linked immunosorbent assay (ELISA) package is opened on the plate
After use, the Flat noodles shall be stored in a sealed bag.
2. There may be crystal precipitation in concentrated washing solution. When diluting, it can be dissolved by heating in a water bath, and washing does not affect the results.
3. Sample dispensers should be used for each step of sample addition, and their accuracy should be regularly checked to avoid experimental errors. The best time to add samples is once
Control within 5 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 OD value)
If the OD value is greater than the first well of the standard well, please dilute the sample diluent by a certain factor (n times) before measuring
Please multiply the final product by the total dilution factor (x n x 5) when calculating.
5. The sealing film is only for one-time use to avoid cross contamination.
6. Please store the substrate away from light.
7. Strictly 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 8. All samples, detergents, and various waste materials should be treated as infectious agents.
9. Components from different batch numbers of this reagent must not be mixed.

Specifications
96 servings/box

Storage conditions and expiration date
1. Storage of reagent kit:; 2-8 ℃.
2. Validity period: 6 months