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This Bosterbio ELISA kit is designed to measure the concentration of Testosterone(T) 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 Testosterone(T) and Rabbit anti- Human Testosterone(T) Antibody to form a coated secondary antibody-anti- Testosterone(T) 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 Testosterone(T) in human serum.
| Product Name | Human Testosterone(T) ELISA Kit (Competitive ELISA) |
|---|---|
| SKU/Catalog Number | EK7005 |
| Description | Competitive High Sensitivity ELISA kit for Quantitative Detection of Human Testosterone(T) 96wells/kit, with removable strips. |
| Cite This Product | Human Testosterone(T) ELISA Kit (Competitive ELISA) (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7005) |
| 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 | 0.1 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 | 0.1-20 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 Testosterone(T) Standards(S0~S5) | 6 | 0.5ml | Testosterone(T) (0, 0.1, 0.4, 1.6, 5, 20 ng/ml), 0.02M PBS, 50% detoxification serum, 0.1% Proclin-300 |
| HRP Conjugated antigen | 1 | 6ml | HRP Conjugated Human Testosterone(T), 0.02M PBS, 2% BSA, 0.01% azophloxine, 0.1% Proclin-300 |
| Antibody | 1 | 6ml | Rabbit anti- Human Testosterone(T) Antibody, 0.02M PBS, 2% BSA, 0.1% Proclin-300, 0.01% Evans Blue |
| Controls | 2 | 0.5ml | Testosterone(T), 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.
Testosterone (17β-hydroxyandrost-4-ene-3-one) is a C19 steroid with an unsaturated bond between C-4 and C-5, a ketone group in C-3 and a hydroxyl group in the β position at C-17. This steroid hormone has a molecular weight of 288.4. Testosterone is the most important androgen secreted into the blood. In males, testosterone is secreted primarily by the Leydig cells of the testes; in females ca. 50% of circulating testosterone is derived from peripheral conversion of androstenedione, ca. 25% from the ovary and ca. 25% from the adrenal glands. Testosterone is responsible for the development of secondary male sex characteristics and its measurements are helpful in evaluating the hypogonadal states. In women, high levels of testosterone are generally found in hirsutism and virilization, polycystic ovaries, ovarian tumors, adrenal tumors and adrenal hyperplasia. In men, high levels of testosterone are associated to the hypothalamic pituitary unit diseases, testicular tumors, congenital adrenal hyperplasia and prostate cancer. Low levels of testosterone can be found in patients with the following diseases: Hypopituitarism, Klinefelter’s syndrome, Testicular feminization, Orchidectomy and Cryptorchidism, enzymatic defects and some autoimmune diseases. The Testosterone EIA kits are designed for the measurement of total Testosterone in human serum.
Click the images to enlarge.
Human Testosterone(T) 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. Collect blood specimens and separate the serum immediately.
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.
5. Do not use grossly lipemic specimens.
6. Please note: Samples containing sodium azide should not be used in the assay.
20X Enzyme conjugate: Prepare 1X working solution at 1:20 with assay diluent (e.g. Add 0.1ml of the Testosterone enzyme conjugate concentrate to 1.9ml of assay diluent).
Wash Buffer: 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).
All reagents should be brought to room temperature (20-25°C) before use.
Before proceeding with the assay, bring all reagents to room temperature (20-25°C).
1. Pipette 25 ?l of the standards, control or specimen into the assigned well.
2. Add 100 ?l of working Testosterone-enzyme conjugate reagent to all wells (see Reagent Preparation Section).
3. Swirl the microplate gently for 20-30 seconds to mix the reagents.
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