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This Bosterbio ELISA kit is designed to measure the concentration of Progesterone(P) 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 Progesterone(P) and Rabbit anti- Human Progesterone(P) Antibody to form a coated secondary antibody-anti- Progesterone(P) antibody- Progesterone(P) (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 Progesterone(P) in human serum.
| Product Name | Human Progesterone(P) ELISA Kit (Competitive ELISA) |
|---|---|
| SKU/Catalog Number | EK7004 |
| Description | Competitive High Sensitivity ELISA kit for Quantitative Detection of Human Progesterone(P). 96wells/kit, with removable strips. |
| Cite This Product | Human Progesterone(P) ELISA Kit (Competitive ELISA) (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7004) |
| 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 | 0.2 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 | 1-30 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 | Goat Anti- Rabbit Secondary Antibody, Polystyrene micro-well plate |
| Human Progesterone(P) Standards(S0~S5) | 6 | 0.5ml | Progesterone(P) (0, 1, 3, 6, 12, 30 ng/ml), 0.02M PBS, 50% detoxification serum, 0.1% Proclin-300 |
| HRP Conjugated antigen | 1 | 6ml | HRP Conjugated Human Progesterone(P), 0.02M PBS, 2% BSA, 0.01% azophloxine, 0.1% Proclin-300 |
| Antibody | 1 | 6ml | Rabbit anti- Human Progesterone(P) Monoclonal Antibody, 0.02M PBS, 2% BSA, 0.1% Proclin-300, 0.01% Evans Blue |
| Controls | 2 | 0.5ml | Progesterone(P), 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.
Progesterone is a C21 steroid which is synthesized from both tissue and circulating cholesterol. Cholesterol is transformed to Progesterone which is then converted via a combined dehydrogenase and isomerase to progesterone. The principle production sites are the adrenals and ovaries and the placenta during pregnancy. The majority of this steroid is metabolized in the liver to pregnanediol and conjugated as a glucuronide prior to excretion by the kidneys. Progesterone exhibits a wide variety of end organ effects. The primary role of progesterone is exhibited by the reproductive organs. In males, progesterone is a necessary intermediate for the production of corticosteroids and androgens. In females, progesterone remains relatively constant throughout the follicular phase of the menstrual cycle. The concentration then increases rapidly following ovulation and remains elevated for 4-6 days and decreases to the initial level 24 hours before the onset of menstruation. In pregnancy, placental progesterone production rises steadily to levels of 10 to 20 times those of the luteal phase peak. Progesterone measurements are thus performed to determine ovulation as well as to characterize luteal phase defects. Monitoring of progesterone therapy and early stage pregnancy evaluations comprise the remaining uses of progesterone assays. Progesterone EIA kits are designed for the measurement of total progesterone in human serum or plasma.
Click the images to enlarge.
Human Progesterone(P) 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. It is recommended to collect serum samples with commercially available equipments. The serum samples should be completely colorless even the slight red color shows blood contamination.
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.
Working Reagent, A Progesterone-enzyme Conjugate Solution: Dilute the Progesterone enzyme conjugate 1:21 with assay diluent in a suitable container. For example, dilute 100?l of conjugate with 2ml of assay diluent buffer for 10 wells (A slight excess of solution is made).
Wash Buffer: Prepare 1X Wash Buffer by adding the contents of the bottle (25ml, 20X) to 475 ml of distilled or deionized water. Store at room temperature.
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 10 ul of Progesterone standards, control and patient’s serum samples.
3. Add 200 ul of Progesterone Enzyme Conjugate to all wells.
4. Incubate for 60 minutes at room temperature (20-25°C).
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