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This Bosterbio ELISA kit is designed to measure the concentration of Estrodial (E2) in human serum by Competitive ELISA (Enzyme-Linked Immunosorbent Assay). A goat anti-rabbit coated microplate is used to prepare a solid phase secondary antibody, and then add samples, HRP Conjugated Human Estrodial (E2) and Rabbit anti- Human Estrodial (E2) Antibody to form a coated secondary antibody-anti- Estrodial (E2) antibody- Estrodial (E2) (HRP) complex. The amount of bound peroxidase conjugate is inversely proportional to the concentration of estradiol in the sample. After color development, the absorbance (O.D. value) is measured by a microplate reader. The concentration-absorbance curve can be fitted by computer or mapping to calculate the concentration of Estrodial (E2) in human serum.
| Product Name | Human Estrodial (E2) ELISA Kit (Competitive ELISA) |
|---|---|
| SKU/Catalog Number | EK7049 |
| Description | Competitive High Sensitivity ELISA kit for Quantitative Detection of Human Estrodial (E2). 96wells/kit, with removable strips. |
| Cite This Product | Human Estrodial (E2) ELISA Kit (Competitive ELISA) (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7049) |
| 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 | 40 pg/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 | 40-1400 pg/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 | Goat Anti- Rabbit Secondary Antibody, Polystyrene micro-well plate |
| Human Estrodial (E2) Standards(S0~S5) | 6 | 0.5ml | Estrodial (E2) (0, 40, 120, 350, 700, 1400 pg/ml), 0.02M PBS, 50% detoxification serum, 0.1% Proclin-300 |
| HRP Conjugated antigen | 1 | 6ml | HRP Conjugated Human Estrodial (E2), 0.02M PBS, 2% BSA, 0.01% azophloxine, 0.1% Proclin-300 |
| Antibody | 1 | 6ml | Rabbit anti- Human Estrodial (E2) Antibody, 0.02M PBS, 2% BSA, 0.1% Proclin-300, 0.01% Evans Blue |
| Controls | 2 | 0.5ml | Estrodial (E2), 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 | HSD17B1 |
|---|---|
| Uniprot ID | P14061 |
| Reseach Areas | Energy Metabolism, Energy Transfer Pathways, Growth Factors, Growth Factors/Hormones, Metabolic Signaling Pathways, Metabolism, Neuroscience, Pathways And Processes, 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 Estrodial (E2) ELISA Kit standard curve
1. For Research Use Only. Not for use in diagnostic procedures.
2. For laboratory use.
3. The chemotherapeutic drug fulvestrant interferes with the assay. Therefore, we recommend not to use this kit for the determination of estradiol levels in patients undergoing treatment with this agent.
4. Potential biohazardous materials: The Standard set contains 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.
5. 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.
6. Do not pipette by mouth. Do not smoke, eat, or drink in the areas in which specimens or kit reagents are handled.
7. The components in this kit are intended for use as an integral unit. The components of different lots should not be mixed.
1. Collect blood specimens and separate the serum immediately.
2. Typically, specimens may be stored refrigerated at (2-8. C) for 5 days. For long term storage 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.
1. 20X Enzyme conjugate: Prepare 1X working solution at 1:20 with assay diluent (e.g. Add 0.1ml of the E2 enzyme conjugate concentrate to 1.9ml of assay diluent)
2. 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. Bring all reagents to room temperature (20-25 oC) before use.
2. Secure the desired number of coated wells in the holder.
3. Dispense 25ul of standards, specimens and controls into appropriate wells.
4. Dispense 100ul of working solution of Estradiol enzyme conjugate into each well.
5. Mix well by placing on shaker for 10-20 seconds
6. Incubate at room temperature (20-25°C) for 60 minutes.
7. Remove liquid from all wells. Wash wells three times with 300ul of 1X wash buffer. Blot on absorbance paper or paper towel.
8. Dispense 100ul of TMB Reagent into each well. Gently mix for 10 seconds.
9. Incubate at room temperature (20-25°C) for 30 minutes.
10. Stop the reaction by adding 50ul of Stop Solution to each well.
11. Gently mix 30 seconds. It is important to make sure that all the blue color changes to yellow color completely.
12. Read absorbance at 450 nm with a microplate reader within 15 minutes.
1. Calculate the mean absorbance value (A450) for each set of reference standards, controls and samples. This standard curve is for the purpose of illustration only, and should not be used to calculate unknowns. against its concentration in pg/ml on a linear-linear graph paper, with absorbance values on the vertical or Y axis, and concentrations on the horizontal or X axis.
3. Use the mean absorbance values for each specimen to determine the corresponding concentration of B2MG in .g/ml from the standard curve.
4. Any values obtained for diluted samples must be further converted by applying the appropriate dilution factor in the calculations.
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