Preparation of rat activin A ELISA kit reagent: 2. Dilution of wash buffer (50×): dilute with distilled water 50 times. Rat Activin A ELISA Kit Procedure: Result judgment and analysis: The Chinese herbal medicine extracts raw material supplier and manufacturer -- Organic Herb Inc is a China-based company which is committed to producing , developing and exploring straight herbal powder ,plant extracts, nutraceutical ingredients, effective components of Chinese herbs and related products for the pharmaceuticals, beverage , health food and cosmetics industries.OHI follow the c-GMPs for herbal raw material and straight powder and we pledge that we will always uphold the highest standards of safety for our customers around the world with competitive market prices. OHI obtains straight powder from organic plant source through the highest standard of manufacture process , so that every one of our powder provides the most nutrients possible. Raw Material / Straight Powder Raw Material,Straight Powder,Ginseng Straight Powder,Extract Powder Raw Materials Organic Herb Inc. , https://www.extracts.pl
1. Standard: The serial dilution of the standard should be prepared during the experiment and cannot be stored. Mix the standard shakes before dilution. The dilution ratio is as follows:
500pg/ml (Standard No. 6) The original concentration is directly added to 50ul without dilution.
250pg/ml (No. 5 standard) 100ul of the original standard is added to 100ul of standard dilution
125pg/ml (Standard No. 4) 100ul of Standard No. 5 is added to 100ul of standard dilution
62.5pg/ml (Standard No. 3) 100ul of Standard No. 4 is added to 100ul of standard dilution
31.2pg/ml (Standard No. 2) 100ul of Standard 3 is added to 100ul of standard dilution
15.6pg/ml (Standard No. 1) 100ul of Standard 2 is added to 100ul of standard dilution
0pg/ml (blank control) The original concentration is directly added to 50ul without dilution.
1. Mix all reagents thoroughly before use. Do not allow the liquid to generate a large amount of foam, so as to avoid adding a large amount of air bubbles during the loading, resulting in errors in the loading.
2. Determine the number of slats required based on the number of samples to be tested plus the number of standards. Multiple holes are recommended for each standard and blank hole. Each sample is determined according to its own quantity, and it is possible to use the double hole as much as possible to make a double hole.
3. Add 50 ul of the diluted standard to the reaction well, and add 50 ul of the sample to be tested to the reaction well. Immediately add 50 ul of biotinylated antibody. Cover the membrane, mix gently by shaking, and incubate for 45 minutes at 37 °C.
4. Remove the liquid from the well, fill each well with the washing solution, shake for 30 seconds, remove the washing solution, and pat dry with absorbent paper. Repeat this operation 4 times. If washing with a washer, the number of washings is increased once.
5. Add 100 ul of affinity streptavidin-HRP to each well, mix gently by shaking, and incubate for 30 minutes at 37 °C.
6. Remove the liquid from the wells, fill each well with the washing solution, shake for 30 seconds, remove the washing solution, and pat dry with absorbent paper. Repeat this operation 4 times. If washing with a washer, the number of washings is increased once.
7. Add 50 ul of each of the substrates A and B to each well, mix gently by shaking, and incubate at 37 ° C for 5 minutes. Avoid lighting.
8. Remove the ELISA plate and quickly add 50 ul of stop solution. Immediately after adding the stop solution, the results should be determined.
9. Determine the OD value of each well at a wavelength of 450 nm.
1. Instrument value: read the OD value of each hole on a microplate reader with a wavelength of 450 nm.
2. The absorbance OD value is the ordinate (Y), and the corresponding HBSAG standard concentration is the abscissa (X), and the corresponding curve is obtained. The HBSAG content of the sample can be converted into the corresponding concentration according to the OD value from the standard curve. Multiply by the dilution factor; or calculate the regression equation of the standard curve using the concentration of the standard and the OD value, substitute the OD value of the sample into the equation, calculate the sample concentration, and multiply by the dilution factor, which is the actual concentration of the sample.
3. Detection range: 0-500pg/ml
4. Sensitivity: 1.95pg/ml
Rat Activin A ELISA Kit Instructions
Rat Activin A ELISA 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 (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 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. In the case of an English manual, the English manual shall prevail.