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The Bosterbio OneStep Human Thyroid-Stimulating Hormone (TSH) Pre-Coated ELISA (Enzyme-Linked Immunosorbent Assay) kit is a solid phase immunoassay specially designed to measure Human Thyroid-Stimulating Hormone (TSH) with a 96-well strip plate that is pre-coated with antibody specific for Thyroid-Stimulating Hormone (TSH). The detection antibody is a HRP conjugated antibody specific for Thyroid-Stimulating Hormone (TSH). The capture antibody is monoclonal antibody from mouse, the detection antibody is monoclonal antibody from mouse. The kit is analytically validated with ready to use reagents. To measure Human Thyroid-Stimulating Hormone (TSH), add standards and samples to the wells, then add the HRP conjugated detection antibody. Wash away the unbounded protein and HRP conjugated detection antibody. TMB is substrate to HRP and will be catalyzed to produce a blue color product, which changes into yellow after adding acidic stop solution. Upon addition of the substrate, the density of the yellow product is linearly propotional to Human Thyroid-Stimulating Hormone (TSH) in the sample. Read the density of the yellow product in each well using a plate reader, and benchmark the sample wells' readings against the standard curve to determine the concentration of Human Thyroid-Stimulating Hormone (TSH) in the sample. For more information on assay principle, protocols, and troubleshooting tips, see Boster's ELISA Resource Center at https://www.bosterbio.com/elisatechnical-resource-center.
| Product Name | Human Thyroid Stimulating Hormone (TSH) OneStep ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7098 |
| Description | Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human Thyroid-Stimulating Hormone (TSH). 96wells/kit, with removable strips. |
| Cite This Product | Human Thyroid Stimulating Hormone (TSH) OneStep ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7098) |
| Validated Species | Human |
| Application | ELISA *Our Boster Guarantee covers the use of this product in the above tested applications. **For positive and negative control design, consult "Tissue specificity" under Protein Target Info. |
| Cross Reactivity | There is no detectable cross-reactivity. |
| Pack Size | 96wells/kit, with removable strips. |
| Sensitivity | 0.10 mIU/L *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 | 0.3-12 mIU/L *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 |
| Anti-Human Thyroid-Stimulating Hormone (TSH) Pre-coated 96-well strip microplate | 1 | 8 strips of 12 wells | Anti- Thyroid-Stimulating Hormone (TSH) monoclonal antibody, Polystyrene micro-well plate |
| Human Thyroid-Stimulating Hormone (TSH) Standards(S1~S6) | 6 | 0.5ml | Thyroid-Stimulating Hormone (TSH) (0.3, 0.6, 1.5, 3.0, 6.0, 12.0mIU/L), 0.02M PBS, 2% BSA, 0.1%Proclin-300 |
| HRP Conjugated anti-Human Thyroid-Stimulating Hormone (TSH) antibody | 1 | 6ml | HRP Conjugated anti-Human Thyroid-Stimulating Hormone (TSH) antibody, 0.02M PBS, 20% new-born calf serum, 0.1%Proclin-300, 0.01% azophloxine |
| Controls | 2 | 1ml | Thyroid-Stimulating Hormone (TSH), 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.
You can check the tissue specificity below for information on selecting positive and negative control.
| Gene Name | TSHR |
|---|---|
| Uniprot ID | P16473 |
| Reseach Areas | Endocrine System, Growth Factors/Hormones, Neuroscience, Signal Transduction| |
*if product is indicated to react with multiple species, protein info is based on the human gene.
Click the images to enlarge.
Human TSH ELISA Kit standard curve
1. For Research Use Only. Not for use in diagnostic procedures.
2. For Laboratory Use.
3. Potential biohazardous materials: The standards set 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, as 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
4. Do not pipette by mouth. Do not smoke, eat, or drink in the areas in which specimens or kit reagents are handled.
5. The components in this kit are intended for use as an integral unit. The components of different lots should not be mixed.
6. It is recommended that standards, control and serum samples be run in duplicate.
7. 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.
1. Collect blood specimens and separate the serum immediately.
2. Specimens may be stored refrigerated at (2-8. 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.
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 (20-25°C).
Prior to assay, allow reagents to stand at room temperature (20-25°C). Gently mix all reagents before use.
1. Place the desired number of coated strips into the holder
2. Pipet 50ul of TSH standards, control and patient specimens into designated wells.
3. Add 100ul of ready to use conjugate reagent to all wells. Shake for (10-30) sec.
4. Cover the plate and incubate for 60 minutes at room temperature (20-25°C), with shaking (at 600rpm).
5. Remove liquid from all wells. Wash wells four times with 300ul of 1X wash buffer. Blot on absorbent paper towels.
6. Add 100ul of TMB substrate to all wells.
7. Incubate for 15 minutes at room temperature (DO NOT Shake).
8. Add 50ul of stop solution to all wells. Shake the plate gently to mix the solution.
9. Read absorbance on ELISA Reader at 450 nm within 15 minutes after adding the stopping solution.
(See Reagent Preparation Section). Blot On Absorbent Paper Towels.
1. Check TSH standard value on each standard vial. This value might vary from lot to lot. Make sure you check the value on every kit. See example of the standard attached.
2. Construct a standard curve by plotting the mean absorbance obtained for each reference (vertical axis) versus the TSH standard concentrations (horizontal axis) on a linear graph paper. Draw the best curve through the points.
3. Read the absorbance for controls and each unknown sample from the curve. Record the value for each control or unknown sample.
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