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The Bosterbio β-hCG Pre-Coated ELISA (Enzyme-Linked Immunosorbent Assay) kit is a solid phase immunoassay specially designed to measure β-hCG with a 96-well strip plate that is pre-coated with antibody specific for β-hCG. The detection antibody is a HRP conjugated antibody specific for β-hCG. 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 β-hCG, 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 β-hCG 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 β-hCG 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 | Beta-human Chorionic Gonadotropin(β-hCG) ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7062 |
| Description | Sandwich High Sensitivity ELISA kit for Quantitative Detection of β-hCG. 96wells/kit, with removable strips. |
| Cite This Product | Beta-human Chorionic Gonadotropin(β-hCG) ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7062) |
| 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 | 2.0 IU/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 | 8-240 IU/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 and Urine *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-hCG-β Pre-coated 96-well strip microplate | 1 | 8 strips of 12 wells | Anti-hCG-β monoclonal antibody, Polystyrene micro-well plate |
| hCG-β Standards(S0~S5) | 6 | 0.5ml | hCG-β (0, 8, 16, 40, 100, 240 IU/L), 0.02M PBS, 20% new-born calf serum, 0.1%Proclin-300 |
| HRP Conjugated anti-hCG-β antibody | 1 | 6ml | HRP Conjugated anti-hCG-β antibody, 0.02M PBS, 20% new-born calf serum, 0.01% azophloxine, 0.1%Proclin-300 |
| Controls | 2 | 0.5ml | hCG-β, 100% natural protein, 0.1%Proclin-300 |
| 10 X Urine Sample Dilute Buffer Concentrate | 1 | 15ml | 0.2M PBS, 20% new-born calf serum, 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.
Click the images to enlarge.
β-hCG ELISA Kit standard curve
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, 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 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, bring all reagents to room temperature (20-25°C). Gently mix all reagents before use.
1. Place the desired number of coated strips into the holder. Replace any unused microwell strips back into the foil pouch, seal, and store at 2-8°C.
2. Pipet 25ul of hCG standards, controls, and samples in to appropriate wells.
3. Add 100ul of Conjugate Reagent to all wells.
4. Incubate for 60 minutes at room temperature (20-25oC).
5. Remove liquid from all wells. Wash wells three 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.
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.
Diluted 1:10 By Adding 50ul Urine To 450ul Sample Diluent. Use Same Assay Procedure As For Serum Test.
1. Standard values may vary slightly with each lot. Be sure to use the correct value (printed on the vial label and included with the Certificate of Analysis. The standard below is only an example.
2. To construct the standard curve, plot the absorbance for the TSH standards versus the hCG standard concentrations in mIU/ml (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.
4. Values above 250 mIU should be retested after diluting with"0" standard.
1. Do not use sodium azide as preservative. Sodium azide inhibits HRP enzyme activities.
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