Plant Protease ELISA Kit

Plant Protease ELISA Kit

Specification: 96T/48T

Purpose: Used for detecting serum, plasma, cell culture supernatant and other samplesThe concentration of plant protease (PROTEASE).

Please read the instruction manual carefully before use. If you have any questions, please contact us

Please refer to the outer packaging label of the reagent kit for the specific shelf life. Please use the reagent kit within its shelf life. Please provide the product item number and production date (see box label) when contacting, so that we can serve you more efficiently.

【 Performance of reagent kit 】

Physical properties: Each liquid component is clear and transparent, without precipitation or flocculent substances. Microporous aluminum foil bags should be vacuum packed without damage or air leakage.

Calibration curve linearity: The correlation coefficient r value of the dose response curve of the calibration sample is greater than or equal to 0.9900.

Precision: The coefficient of variation (CV%) within the batch is less than 10%; The coefficient of variation (CV%) between batches is less than 15%.

Recovery rate: The recovery rate is between 85% and 115%.

Sensitivity: This kit recognizes plant protease (PROTEASE) and has no cross reactivity with structurally similar compounds.

Stability: Stored at 2 ℃ -8 ℃, with a shelf life of 6 months.

【 Experimental Principle 】

Plant Protease ELISA KitApplication of double antigen sandwich method to determine the expression of indicators in specimens. Encapsulate the purified antigen onto a microplate to produce a solid-phase antigen that can bind to the indicators in the sample. After thorough washing, add the substrate TMB for color development. TMB is converted to blue under the catalysis of HRP enzyme and to the final yellow under the action of acid. The absorbance (OD value) measured by enzyme-linked immunosorbent assay (ELISA) at a wavelength of 450nm is positively correlated with the concentration of plant protease (PROTEASE) in the test sample. Fitting the calibration curve can calculate the concentration in the sampleThe concentration of plant protease (PROTEASE).

【 Kit Composition 】

Note:

1. Before use, please check if the labels and quantities of the reagents in the kit match the table.

2. The reagent kit should be stored at 2-8 ℃ and expired kits should not be used.

3. If the microporous plate is not used up in a single use, remember to seal it and store it at 2-8 ℃.

4. If the components of the reagent kit need to be reused, please ensure that they have not been contaminated after the last use.

Preparation and Preservation of Samples

The following is just a general guide for sample collection and preservation. Sodium azide should not be used as a preservative during the collection and storage of all samples. If the sample is not analyzed immediately, it should be packaged and frozen for storage, and repeated freeze-thaw cycles should be avoided.

Cell culture supernatant: Centrifuge to remove sediment, immediately analyze or package and freeze at -20 ° C.

Serum: Collect blood in a clean test tube, coagulate at room temperature for 30 minutes, centrifuge at 2000 × g for 20 minutes, and collect serum. Immediately analyze or package and freeze at -20 ℃ for storage.

Plasma: Anticoagulated with heparin, citrate, or EDTA, centrifuged at 2-8 ℃ for 20 minutes within 30 minutes after drawing. To eliminate the influence of platelets, it is recommended to further centrifuge 10000 × g at 2-8 ℃ for 10 minutes. Immediately analyze or freeze at -20 ℃.

Cell lysate: For adherent cells, remove the culture medium and wash once with PBS, physiological saline, or serum-free culture medium. Add an appropriate amount of lysis buffer and blow it several times with a pipette to ensure full contact between the lysis buffer and the cells. Usually, after 10 seconds, the cells will be lysed. For suspended cells, collect them by centrifugation and wash them once with PBS, physiological saline, or serum-free culture medium. Add an appropriate amount of lysis buffer, use a pipette to blow and disperse the cells, and lightly tap with your fingers to fully lyse the cells. After sufficient lysis, centrifuge at 1000014000 × g for 3-5 minutes and collect the supernatant. Immediately analyze or package and freeze at -20 ℃ for storage.

Urine: Collect with a sterile tube and centrifuge at 2000 × g for 20 minutes. Carefully collect the supernatant. If precipitation forms, it should be centrifuged again.

【 Precautions 】

1. This kit is only for in vitro research use and is not intended for clinical diagnosis.

2. Please wear experimental clothing and latex gloves for protection during the experiment. Especially when testing blood or other bodily fluid samples, please follow the national biological laboratory safety protection regulations.

3. Strictly follow the prescribed time and temperature for incubation to ensure accurate results. All reagents must reach room temperature of 20-25 ℃ before use. Immediately refrigerate and store the reagents after use.

4. Incorrect board washing may result in inaccurate results. Ensure to absorb as much liquid as possible from the well before adding the substrate. Do not let the micropores dry out during the incubation process.

5. Eliminate residual liquid and fingerprints at the bottom of the board, otherwise it will affect the OD value.

6. The substrate color solution should be colorless or very light in color.

7. Avoid cross contamination between reagents and specimens to prevent erroneous results.

8. Avoid direct exposure to strong light during storage and incubation.

9. After balancing to room temperature, open the sealing bag to prevent water droplets from condensing on the cold Flat noodles.

10. Any reaction reagent should not come into contact with bleach solvents or the strong gases emitted by bleach solvents. Any bleaching component will destroy the biological activity of the reactants in the reagent kit.

11. The enzyme-linked immunosorbent assay (ELISA) used for detection requires the installation of a filter capable of detecting wavelengths of 450 ± 10nm,

The optical density range is between 0-3.5. It is recommended to preheat 15 minutes before use.

12. Do not mix or replace the reagents in this kit with reagents from other batch numbers or sources.

13. The EP tube and suction head used in the experiment are disposable and must not be mixed.

14. Do not use expired reagents.

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