SKU: 30973258600

Rat TSH ELISA Kit

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Description

Rat TSH ELISA KitProduct Specification Usage Sample Processing and Requirements 1. Serum: Place the whole blood specimen collected in a serum separator tube at room temperature for 2 hours or at 4C overnight, then centrifuge at 1000g for 20 minutes. Remove the supernatant and store it at 20C or 80C. However, repeated freezing and thawing should be avoided. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Within 30 minutes of collection,

Product Specification

Usage

Sample Processing and Requirements

1.
Serum: Place the whole blood specimen collected in a serum separator tube at room temperature for 2 hours or at 4°C overnight, then centrifuge at 1000×g for 20 minutes.
Remove the supernatant and store it at -20°C or -80°C.
However, repeated freezing and thawing should be avoided.

2.
Plasma: Collect the specimen using EDTA or heparin as an anticoagulant.
Within 30 minutes of collection, centrifuge at 1000×g for 15 minutes at 2-8°C.
Remove the supernatant and test it.
Alternatively, store it at -20°C or -80°C.
However, repeated freezing and thawing should be avoided.

3.
Tissue Homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove any residual blood (lysed red blood cells in the homogenate will affect the measurement results).
Weigh the tissue and mince it.
Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL of PBS.
The specific volume can be adjusted according to experimental needs and recorded.
It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice.
To further lyse tissue cells, the homogenate can be sonicated or repeatedly freeze-thawed.
Finally, centrifuge the homogenate at 5000×g for 5-10 minutes.
Remove the supernatant for analysis.

4.
Cell culture supernatant or other biological specimens: Centrifuge at 1000×g for 20 minutes.
Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing.
Note: Hemolysis of the specimen will affect the final test results, so hemolyzed specimens are not suitable for this test.

Reagent Preparation
After removing the kit from the refrigerator, allow it to equilibrate to room temperature before use.
Dilution of 20× Wash Buffer: Dilute 1:20 with distilled water, i.e., add 1 part 20× Wash Buffer to 19 parts distilled water.

Procedure
1.
Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 20 minutes.
Seal the remaining strips in a ziplock bag and return them to 4°C.

2.
Set up standard wells and sample wells.
Add 50 μL of standard solution of varying concentrations to each standard well.

3.
Add 50 μL of the sample to be tested to the sample wells; leave blank wells untouched.

4.
Add 100 μL of horseradish peroxidase (HRP)-labeled detection antibody to each standard and sample well, except for the blank well.
Seal the wells with sealing film and incubate at 37°C in a waterbath or incubator for 60 minutes.

5.
Discard the liquid, pat dry on absorbent paper, and fill each well with wash buffer (350 μL).
Let stand for 1 minute, then discard the wash buffer and pat dry on absorbent paper.
Repeat this process five times (a plate washer can also be used).

6.
Add 50 μL each of substrates A and B to each well and incubate at 37°C in the dark for 15 minutes.

7.
Add 50 μL of stop solution to each well.
Within 15 minutes, measure the OD value of each well at a wavelength of 450 nm.

Calculation of Experimental Results:
Using the OD value of the measured standard as the horizontal axis and the concentration of the standard as the vertical axis, plot a standard curve on graph paper or using relevant software.
Obtain a linear regression equation.
Substitute the OD value of the sample into the equation to calculate the sample concentration.
This figure is for reference only: Standard Curve.

Sensitivity The minimum detection concentration is less than 0.1 mU/L.
Species Reactivity Rat
Theory The kit uses a double-antibody one-step sandwich enzyme-linked immunosorbent assay (ELISA). Samples, standards, and HRP-labeled detection antibodies are added sequentially to microwells pre-coated with a rat thyroid-stimulating hormone (TSH) capture antibody. The assay is incubated and thoroughly washed. The color is developed using the substrate TMB, which converts to blue under the catalysis of peroxidase and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of rat thyroid-stimulating hormone (TSH) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration.
Detection Type Used for in vitro quantitative detection of rat thyroid-stimulating hormone (TSH) content in serum, plasma, tissue homogenate and related liquid samples.
Composition

Name

96 T

Remarks

Microwell enzyme plate

12 holes×8 strips

None

Standard product

0.3mL*6 tube

None

Sample diluent

6mL

None

Detection antibody-HRP

10mL

None

20×Wash Buffer

25mL

Dilute according to instructions

Substrate A

6mL

None

Substrate B

6mL

None

Stop solution

6mL

None

Sealing film

2 Zhang

None

Remarks:

1. The concentrations of the standard substances are: 20, 10, 5, 2.5, 1.25, and 0.625 mU/L.

2. After testing a large number of normal specimens, the normal concentration values of the specimens are all within the detection range provided by the kit. During the experiment, 50 μL of sample can be directly sampled. If some sample values exceed the maximum standard concentration, the sample can be appropriately diluted with sample diluent before the experiment.

General Notes

1. Strictly follow the specified incubation time and temperature to ensure accurate results. All reagents must reach room temperature (20-25°C) before use. Refrigerate reagents immediately after use.

2. Improper plate washing can lead to inaccurate results. Ensure that the liquid in the wells is as dry as possible before adding substrate. Do not allow the microwells to dry out during incubation.

3. Remove any residual liquid and fingerprints from the bottom of the plate, otherwise it will affect the OD value.

4. The substrate developer solution should be colorless or very light in color. Substrate solution that has turned blue should not be used.

5. Avoid cross-contamination of reagents and specimens to prevent erroneous results.

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

7. Allow the sealed bag to equilibrate to room temperature before opening to prevent water droplets from condensing on the cold plate strips.

8. No reaction reagents should come into contact with bleaching solvents or the strong fumes emitted by bleaching solvents.

Any bleaching component will destroy the biological activity of the reagents in the kit.

9. Do not use expired products.

10. If there is a possibility of disease transmission, all samples should be managed properly and the samples and detection devices should be handled according to the prescribed procedures.

Storage Temp. Unopened test kit, stored at 2-8°C, has a shelf life of 6 months.
Test Range 0.625 mU/L – 20 mU/L.
Shipping Notes
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Exchange/Return Notes
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SKU: 30973258600

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