Technical guidance for Serpina3g ELISA detection kit for human serine protease inhibitor peptidase inhibitor evolutionary branch 3G

1、 Specification parameters of Serpina3g ELISA detection kit for human serine protease inhibitor peptidase inhibitor evolutionary branch 3G

Chinese name:Serpina3g ELISA detection kit for human serine protease inhibitor peptidase inhibitor evolutionary branch 3G

Specification: 48T 96T

Species: Human

Brand:Zhenke Biology

Detection wavelength: 450nm

Required sample volume: insurance volume 50ul

Scope of application: For scientific research only, not for clinical diagnosis. Clinical specimens can be used for research purposes.

Detection purpose: To determine the content of Serpina3g, a serine protease inhibitor and peptidase inhibitor evolutionary branch, in human serum, plasma, and related liquid samples.

Test sample types: serum, plasma, urine, tissue homogenate, cell supernatant, cerebrospinal fluid, lavage fluid, feces, and other samples

Zhenke supplies ELISA kits for humans, rats, mice, guinea pigs, humans, pigs and dogs, cattle and sheep, chickens and ducks, plants, fish, insects, etc

2、 Composition of ELISA kit

3、 Sample processing and requirements:

1. serumBlood naturally coagulates at room temperature for 10-20 minutes, then centrifuges for about 20 minutes (2000-3000 revolutions per minute). Carefully collect it

If precipitation occurs during storage, it should be centrifuged again.

2. plasmaEDTA or sodium citrate should be selected as anticoagulants according to the requirements of the specimen. After mixing for 10-20 minutes, centrifuge

20 About minutes (2000-3000 revolutions per minute). Carefully collect the supernatant. If sediment forms during storage, it should be collected again

Centrifuge.

3. urineCollect with sterile tubes and centrifuge for about 20 minutes (2000-3000 rpm). Carefully collect the supernatant and store it during the process

If precipitation forms, it should be centrifuged again. Refer to the implementation for pleural effusion, ascites, and cerebrospinal fluid.

4. Cell culture supernatantWhen detecting secretory components, collect them using sterile tubes. Centrifuge for about 20 minutes (2000-3000 rpm)/

Divided). Carefully collect the supernatant. When detecting the components inside cells, dilute the cell suspension with PBS (pH 7.2-7.4), and the cells

The concentration reaches around 1 million/ml. By repeatedly freezing and thawing, cells are destroyed and intracellular components are released. Centrifuge for 20 minutes

Around the clock (2000-3000 revolutions per minute). Carefully collect the supernatant. If precipitation forms during storage, it should be centrifuged again.

5. Organizational specimenAfter cutting the specimen, weigh it. Add a certain amount of PBS, pH 7.4. Quickly freeze and store with liquid nitrogen for preparation

Use it. The specimen remains at a temperature of 2-8 ℃ even after melting. Add a certain amount of PBS (pH 7.4) by hand or homogenizer

Homogenize the specimen thoroughly. Centrifuge for about 20 minutes (2000-3000 rpm). Carefully collect the supernatant. After packaging, one portion is to be prepared

Test and freeze the rest for future use.

6. Extract the specimen as soon as possible after collection, according to relevant literature, and conduct experiments as soon as possible after extraction. If we can't do it right away

For the experiment, the specimen can be stored at -20 ℃, but repeated freezing and thawing should be avoided

7. Samples containing NaN3 cannot be detected because NaN3 inhibits the activity of horseradish peroxidase (HRP).

4、 Attention:

1The 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.

2Concentrated washing solution may precipitate crystals, and can be diluted by heating in a water bath to aid dissolution. Washing 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. The best time for adding 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.

4Please make a standard curve at the same time as each measurement, so that it is easier to make a duplicate 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

When calculating, please multiply it by the total dilution factor (x n x 5).

5The 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.

10. If there is any discrepancy with the English manual, the English manual shall prevail.

5ELISACalculation of reagent kit

Using the concentration of the standard substance as the horizontal axis and the OD value as the vertical axis,

Draw a standard curve on coordinate paper based on the OD of the sample

Find the corresponding concentration from the standard curve; Multiply by dilution again

multiple; Or calculate the standard using the concentration and OD value of the standard substance

The linear regression equation of the quasi curve calculates the OD value of the sample

Substitute into the equation, calculate the sample concentration, and then multiply it by the dilution

The multiple is the actual concentration of the sample.

6、 ELISA kit performance

1.The correlation coefficient R between sample linear regression and expected concentration is above 0.990.

2.Within batch and in batch should be less than 9% and 11%, respectively

3.Storage conditions and expiration date: refrigerated at 2-8 ℃, with a shelf life of 6 months for the reagent kit.

7、 Special Reminder

Due to the current conditions and scientific and technological level, it is not yet possible to comprehensively identify and analyze all raw materials provided by all suppliers, and there may be certain quality and technical risks associated with this product.

1. The final experimental results are closely related to the effectiveness of the reagents, the relevant operations of the experimenter, and the experimental environment at that time. Please be sure to prepare sufficient specimen backups.
2. There may be slight differences in the detection limit, sensitivity, and color development time of the same product from different batches. Please refer to the instructions in the reagent kit for experimental operations. The electronic version of the instructions is for reference only.
3. Only by using all the reagents in this kit can the detection effect be guaranteed, and products from other manufacturers cannot be mixed. Only by strictly following the experimental instructions of this kit can we obtain * test results.
4. Our company is only responsible for the reagent kit itself and is not responsible for the sample consumption caused by the use of the kit. Users are advised to fully consider the possible usage of the sample and reserve sufficient samples before use.
5. The preparation of tissue homogenates or cell extracts using chemical lysates may result in bias in ELISA experimental results due to the introduction of certain chemicals.
6. If the sample is a cell culture supernatant, there may be many interfering factors such as cell status, cell quantity, sampling time, etc., so there may be situations where it cannot be detected.
7. Some natural or recombinant proteins, including prokaryotic and eukaryotic recombinant proteins, may not be detected due to mismatches with the detection and capture antibodies used in this product.