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The Bosterbio OneStep ELISA kit is a solid phase direct ELISA sandwich kit. Instead of adding samples detection antibody and ABC-HRP separately, The OneStep ELISA kit allows the user to add standards, samples and controls to wells in one step, along with the incubation buffer. After a simple washing step, an enzyme conjugate reagent is added into each well. After the excess enzyme conjugate is washed out, the substrate is added into each well. The enzyme catalyzes the substrate yielding a blue color (Amax = 370nm and 652nm) that changes to yellow (Amax = 450nm) upon addition of a sulfuric or phosphoric acid stop solution. The intensity of color developed is directly proportional to the concentration of target protein in the samples. A standard curve is generated relating color intensity to the concentration of target protein.
| Product Name | Human Troponin I OneStep ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7092 |
| Description | Human Troponin I OneStep ELISA Kit, tested with Serum and Plasma. Format: 96wells/kit, with removable strips. |
| Cite This Product | Human Troponin I OneStep ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7092) |
| 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 | 2 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 | 2-75 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 and Plasma *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 |
| 1. Microwells coated with mouse Anti-Tnl | 12x8x1 |
| 2. Reference Standard Set | 1 ml |
| 3. cTnI Enzyme Conjugate Reagent | 13 ml |
| 4. TMB Reagent | 11 ml |
| 5. Stop Solution | 11 ml |
| 6. Wash Concentrate 20x: 1 Bottle | 25 ml |
1. Distilled or deionized water
2. Precision pipettes
3. Disposable pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
4. ELISA reader capable of reading absorbance at 450nm
5. Absorbance paper or paper towel
6. Graph paper
You can check the tissue specificity below for information on selecting positive and negative control.
| Gene Name | Tnni3 |
|---|---|
| Uniprot ID | P48787 |
*if product is indicated to react with multiple species, protein info is based on the human gene.
1. For Research Use Only. Not for use in diagnostic procedures.
2. For Laboratory Use.
3. 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.
4. Optimal results will be obtained by strict adherence to the test protocol. Precise pipetting as well as following the exact time and temperature requirements is essential.
5. Do not pipette by mouth. Do not smoke, eat, or drink in the areas in which specimens or kit reagents are handled.
6. The components in this kit are intended for use as an integral unit. The components of different lots should not be mixed.
7. Control sera and sample diluent contain preserved with sodium azide. Sodium azide may react with lead and copper plumbing to form explosive metal azide. On disposal, flush with a large volume of water.
1. The use of SERUM samples is required for this test.
2. Specimens should be collected using standard venipuncture techniques. Remove serum from the coagulated or packed cells within 60 minutes after collection.
3. Specimens which cannot be assayed within 24 hours of collection should be frozen at 20°C or lower, and will be stable for up to six months.
4. Avoid grossly hemolytic (bright red), lipemic (milky), or turbid samples (after centrifugation).
5. Specimens should not be repeatedly frozen and thawed prior to testing. DO NOT store in "frost free" freezers, which may cause occasional thawing. Specimens which have been frozen, and those which are turbid and/or contain particulate matter, must be centrifuged prior to use.
1. All reagents should be allowed to reach room temperature (20-25°C) before use.
2. Reconstitute each lyophilized standard with 1.0 ml distilled water. Allow the reconstituted material to stand for at least 20 minutes and mix gently. The Reconstituted standards will be stable for up to 8 hours when stored sealed at 2-8°C. Discard the reconstituted Standards after 8 hours. To assure maximum stability of the reconstituted Standards, they should be aliquoted and frozen (.20°C or below) immediately after reconstitution has been achieved. Each aliquoted Standard should be frozen and thawed only once.
3. 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).
1. Secure the desired number of coated wells in holder.
2. Dispense 100 ul of standards, specimens, and controls into appropriate wells.
3. Dispense 100 ul of Enzyme Conjugate Reagent into each well.
4. Thoroughly mix for 30 seconds. It is very important to mix completely.
5. Incubate at room temperature (20-25°C) for 90 minutes.
6. Remove the incubate mixture by flicking plate contents into a waste container.
7. Remove liquid from all wells. Wash wells three times with 300 ul of 1X wash buffer. Blot on absorbance paper or paper towel.
8. Strike the wells sharply onto absorbent paper or paper towels to remove all residual water droplets.
9. Dispense 100 ul of TMB Reagent into each well. Gently mix for 5 seconds.
10. Incubate at room temperature for 20 minutes.
11. Stop the reaction by adding 100 ul of Stop Solution to each well.
12. Gently mix for 30 seconds. It is important to make sure that all the blue color changes to yellow color completely.
13. Read absorbance at 450nm with a microtiter well reader within 15 minutes.
1. Calculate the mean absorbance value (OD450) for each set of reference standards, controls and samples. and standard curve in each experiment. concentration in ng/ml on graph paper, with absorbance on the vertical (y) axis and concentration on the horizontal (x) axis.
3. Using the mean absorbance value for each sample, determine the corresponding concentration of Troponin I (ng/ml) from the standard curve. Depending on experience and/or the availability of computer capability, other methods of data reduction may be employed.
4. Patient samples with cTnI concentrations greater than 100 ng/ml should be diluted 10-fold with vender's Troponin I Sample Diluent. The final cTnI results in ng/mI.
A typical absorbance data and the resulting standard curve from human Chromogranin A EUSA are understanding of the package insert instructions and with adherence to good laboratory practice.
2. Serum samples demonstrating gross lipemia, gross hemolysis, or turbidity should not be used with this test.
3. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
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