Thyroxine(T4)ELISA Test Kit (Serum)

INTENDED USE

Detection Kit for thyroxine(T₄)(Enzyme-Linked ImmunoSorbent Assay , ELISA). It is used in quantitative tests for thyroxine(T4) in human serum.

SUMMARY AND CLINICAL SIGNIFICANCE

The thyroid gland essential for the development of most tissues and for the adjustment of their metabolism. These processes are under the control of thyroid hormones, T4 and T3.

Under stimulation by pituitary thyrotropin (TSH), the thyroid gland essentially secretes T4 with minute amounts of T3. Tissue receptors to thyroid hormones are T3 receptors; T3 represents thus the active entity of thyroid secretions. Nevertheless, T4 determination is of great clinical significance, since serum T4 level is correlated to the secretory rate of the gland. More over T4 represents a precursor hormone available for peripheral deiodination into T3. In assuming serum T4 level, one has to take into consideration both the thyroid secretory rate and deiodination process

PRINCIPLE

T4 kit is based on competitive method. Coat micro well plates with anti-T4 to form solid-phase antibody. The T4 of sample and biotin-T4 compet with anti-T4 on solid-phase. Equilibrium, avidin-HRP added, solid phase antibody-biotinylated T4-avidin-HRP complex was formed. Add substrate and detect absorbent value. Calculate T4 content of patient serum through computer or plotting fitting concentration-absorbent value curve.

MAIN COMPONENTS

Coated Microplate: 1 plate (8×12 wells), Ready to use. Coated with anti-T4 antibody and sealed in an aluminum bag. Remove the strips in the resealable bag with a desiccant to protect from moisture after opened. Store at 2~8℃ until expiration date.

• Biotin-T4: 1 vial of 6ml, Ready to use. Store at 2~8℃ until expiration date.
• Avidin-HRP Conjugate: 1 vial of 6ml, Ready to use. Store at 2~8℃ until expiration date.
• Calibrator: 6 vials of 1ml, Ready to use. Labeled with S₀ to S₅ and the concentration of T4 is 0,20,40, 80,160,320 ng/ml. Store at 2~8℃ until expiration date.
• Control: 2 vials of 1ml, Ready to use. Labeled with high value and low value. Store at 2~8℃ until expiration date. The concentration of low value is 44.3~82.3 ng/ml. The concentration of high value is 112~208 ng/ml.
• Chromogen A: 1 vial of 7ml, Ready to use. Store at 2~8℃ until expiration date.
• Chromogen B: 1 vial of 7ml, Ready to use. Store at 2~8℃ until expiration date.
• Stop Solution: 1 vial of 7ml, Ready to use. Store at 2~8℃ until expiration date.
• Wash buffer: 1 vial of 15ml, Concentrate 20-fold, diluting with deionized water before the assay. Store at 2~8℃ until expiration date.
• Plate sealer: 2 pieces.
• Plastic resealable bag: 1 set.
• Instruction manual: 1 copy.

RELATED TIPS

• This kit is designated for In-Vitro Diagnostic Use Only.
• Wash procedure. Incomplete washing will adversely affect the test results. Wash each well 3 times with about 0.3ml wash buffer. If no automatic washer is available, washing can be performed manually as follows: Invert the plate vigorously to get all wash buffer out and block the rim of well on absorbent paper for a few seconds. Filling each well with water and remain 10 seconds. Repeat these steps 3 times. Blot dry the plate by inverting the plate onto absorbent tissue, and striking a hard surface several times.
• Drip procedure. Mix the bottle gently before use. Violent surge may cause too much foam. Invert the bottle and squeeze one or two drop on absorbent tissue to make sure there is no foam. Take the bottle upright the well and make sure the drop does not touch the rim of wells.
• Read procedure. Using the blank well to correct the zero point of reader if single wavelength reader is used. If double wavelength readers with 450nm and 630nm are used, there is no need to correct the zero point.

Storage. The whole kit should be stored at 2~8℃ for one year. Microplate should be

PREPARATIONS

1. Allow all specimens and reagents to reach room temperature (~25°C) and mix thoroughly by gentle inversion before use.

2.Prepare Wash Buffer by diluting Wash Concentrate 20-fold with deionized water. The diluted wash solution is stable in room temperature for at least one week.

ATTENTIONS

1. Do not exchange reagents from different lots or use reagents from other commercially available kits. The components of the kit are precisely matched for optimal performance of the tests.

2. Specimens with Hyperlipidemia and icterus show no effects on the detection results. But specimen with severe hemolysis will affect the color of the result, other test methods are suggested to use.

3. Allow the reagents and specimens to reach room temperature before use. Shake reagent gently before use.

4. Concentrated washing liquid will produce crystal at room temperature, should be diluted completely before use.

5. Never reuse microplate sealing membrane.

6. Samples containing anti-sperm antibody, anti-endometrial antibody will not affect the results.

7. Each component of the kit was inactivated, however, it should be regarded as potentially infectious.

PRECAUTION FOR USERS

1. Handling should preclude any pipetting by mouth. Use only pipettes with disposable tips for each specimen.

2.Do not mix materials from different master lots. Do not use kit components beyond the expiration date.

3.All materials should be brought to room temperature before use.

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