0
This Bosterbio ELISA kit is designed to measure the concentration of T4 in human serum by Competitive ELISA (Enzyme-Linked Immunosorbent Assay). The polystyrene micro-well plate in this kit has been pre-coated with an anti-Human T4 antibody. Sample or standards are added to the wells along with a fixed quantity of biotinylated T4 and incubated. The T4 found in the sample or standards competes with the biotinylated T4 for limited binding sites on the immobilized anti-Human T4 antibody. Excess unbound biotinylated T4 and sample or standard T4 is washed from the plate. Avidin-HRP conjugate is added, incubated and washed. An enzymatic reaction is then produced through the addition of substrate which is catalyzed by the immobilized HRP to generate a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration is measured by reading the absorbance at 450 nm which is quantitatively proportional to the amount of biotinylated T4 captured in the well and inversely proportional to the amount of T4 which was contained in the sample or standard.
| Product Name | Human Thyroxine (T4) ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7012 |
| Description | Competitive High Sensitivity ELISA kit for Quantitative Detection of Human T4. 96wells/kit, with removable strips. |
| Cite This Product | Human Thyroxine (T4) ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7012) |
| Validated Species | Human |
| Application | ELISA *Our Boster Guarantee covers the use of this product in the above tested applications. |
| Cross Reactivity | There is no detectable cross-reactivity. |
| Pack Size | 96wells/kit, with removable strips. |
| Sensitivity | 15 ng/mL *Sensitivity, or Lower Limit of Detection (LLD), is the minimum level of target protein the ELISA assay can detect. We measure 20 blank wells and if the O.D. value is 2 standard deviations higher than the blanks' average O.D. the sample can be deemed positive. |
|---|---|
| Assay Range | 20-320 ng/mL *This assay range is determined using common samples. For samples with low target protein concentrations, users can adjust the standard curve to extend the lower limit of assay range. |
| Sample Type | Serum *The above listed samples are the ones valided with the assay. If you do not see your sample of interest listed, as long as there is enough level of target protein present in the sample, this Picokine? ELISA kit should detect it. **For protocol and tips regarding preparing your sample of interest, please check our ELISA sample preparation guide. |
| Storage | Store the kit at 2°C to 8°C. Keep microwells sealed in a dry bag with desiccants. The reagents are stable until expiration of the kit. Do not expose reagent to heat, sun, or strong light. Avoid multiple freeze-thaw cycles(Shipped with wet ice.) |
| Description | Quantity | Volume | Buffers |
| Pre-coated 96-well strip microplate | 1 | 8 strips of 12 wells | Rabbit Anti- T4 Antibody, Polystyrene micro-well plate |
| Human T4 Standards(S1~S5) | 5 | 1ml | T4 (20, 40, 80, 160, 320 ng/mL), 100% detoxification serum, 0.1% Proclin-300 |
| Avidin-HRP conjugate | 1 | 6ml | Avidin-HRP conjugate, 0.02M PBS, 20% new-born calf serum, 0.01% azophloxine, 0.1% Proclin-300 |
| Biotinylated antigen | 1 | 6ml | Biotinylated T4, 0.02M PBS, 20% new-born calf serum, 0.1% Proclin-300 |
| Controls | 2 | 1ml | T4, 100% natural protein, 0.1% Proclin-300 |
| 20X Wash Buffer Concentrate | 1 | 15ml | 0.2M PBS containing 0.5% tween 20 |
| Color Developing Reagent A | 1 | 7ml | 11m mol/L Urea hydrogen peroxide |
| Color Developing Reagent B | 1 | 7ml | 2m mol/L 3,3'5,5’-Tetramethylbenzidine |
| Stop Solution | 1 | 7ml | 2mol/L Sulphuric acid |
| Plate Sealers | 2 | Piece |
1. Microplate Reader capable of reading absorbance at 450nm.
2. Automated plate washer (optional)
3. Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples.
4. Deionized or distilledwater.
5. 500ml graduated cylinders.
6. Test tubes for dilution.
T4 is a useful marker for the diagnosis of hypothyroidism and hyperthyroidism. The level of T4 is decreased in hypothyroid patients and is increased in hyperthyroid patients. The level of T4 is normal in Euthyroid individuals.
Click the images to enlarge.
Human T4 ELISA Kit standard curve
1. For Research Use Only. Not for use in diagnostic procedures.
2. Potential biohazardous materials: The calibrator and controls contain human source components, which have been tested and found non-reactive for hepatitis B surface antigen as well as HIV antibody with FDA licensed reagents. However, there is no test method that can offer complete assurance that HIV, Hepatitis B virus or other infectious agents are absent. These reagents should be handled at the Biosafety Level 2, as recommended in the Centers for Disease Control/National Institutes of Health manual, "Biosafety in Microbiological and Biomedical Laboratories" 1984.
3. Do not pipette by mouth. Do not smoke, eat, or drink in the areas in which specimens or kit reagents are handled.
4. The components in this kit are intended for use as an integral unit. The components of different lots should not be mixed.
5. It is recommended that standards, control and serum samples be run in duplicate.
6. Optimal results will be obtained by strict adherence to this protocol. Accurate and precise pipetting, as well as following the exact time and temperature requirements prescribed are essential. Any deviation from this may yield invalid data.
7. Do not use sodium azide as preservative. Sodium azide inhibits HRP enzyme activities.
1. Collect blood specimens and separate the serum immediately.
2. Typically, specimens may be stored refrigerated at (2°C to 8°C) for 5 days. If storage time exceeds 5 days, store frozen at (-20°C) for up to one month.
3. Avoid multiple freeze-thaw cycles.
4. Prior to assay, frozen sera should be completely thawed and mixed well.
5. Do not use grossly lipemic specimens.
T4-enzyme Conjugate Solution: Dilute the T4-enzyme conjugate 1:11 with assay diluent in a suitable container. For example, dilute 80?l of enzyme conjugate with 0.8ml of assay diluent for 16 wells (A slight excess of solution is made). This reagent should be used within twenty-four hours for maximum performance of the assay. Store at 2-8°C).
General Formula:
Amount of Buffer required = Number of wells * 0.05
Quantity of Enzyme conjugate solution necessary = # of wells * 0.005
i.e. = 16 x 0.05 = 0.8ml for Total Conjugate Buffer
16 x 0.005 = 0.08ml (80?l) for enzyme conjugate solution.
Wash Buffer: Prepare 1X Wash buffer by adding the contents of the bottle (25 ml, 20X) to 475 ml of distilled or deionized water. Store at room temperature (18-26°C).
Before proceeding with the assay, bring all reagents, serum references and controls to room temperature (20-25°C).
1. Format the microplates’ wells for each serum reference, control and patient specimen to be assayed in duplicate. Replace any unused microwell strips back into the aluminum bag, seal and store at 2-8°C.
2. Pipette 25 ul of the standards, control or specimen into the assigned well.
3. Add 50 ul of the working T4-enzyme conjugate solution to all wells (see Reagent Preparation Section).
4. Add 50 ul of T4-Antibody-Biotin Solution to all wells.
5. Swirl the microplate gently for 20-30 seconds to mix the reagents.
温馨提示:因厂家更改产品包装、产地或者更换随机附件等没有任何提前通知，且每位咨询者购买情况、提问时间等不同，为此以下回复仅对提问者3天内有效，其他网友仅供参考！若由此给您带来不便请多多谅解，谢谢！
服务热线
0771-3293894
新浪微博
腾讯微博
QQ空间
客服
