>
Antigen, Antibodies, ELISA, Western Blot >
Immunoassay Kits >
ELISA Kit >
Testosterone Rat/Mouse ELISA KitBrand |
Leading Biology | Catalog Number |
EK0001L |
Product Type |
ELISA Kit | Field of Research |
|
Product Overview |
Testosterone is a steroid hormone from the androgen group synthesized by the Leydig cells in the testes in males, the ovaries in females, and adrenal glands in both sexes. It exerts a wide-ranging influence over sexual behaviour, muscle mass and strength, energy, cardiovascular health and bone integrity.
Testosterone biosynthesis coincides with the spermatogenesis and fetal Leydig cell differentiation in the male rat. Several in vivo models including hormone-suppression, hormone-restoration and hypophysectomy were established for the study of the hormonal regulation of spermatogenesis by testosterone (1-3).
In the Brown Norway rat, serum testosterone levels decrease with aging, accompanied by increases in serum FSH. The capacity of Leydig cells to produce testosterone is higher in young than in old rats (4). Testosterone secreted during late gestational and neonatal periods causes significant brain sexual dimorphism in the rat. This results in both sex-specific
behaviour and endocrinology in adults (5).
Analyses concerning the regulation of synthesis reveal that testosterone is able to regulate its own synthesis and indicate that this autoregulation is the result of rapid, specific inhibition by testosterone of 17 alpha-hydroxylase activity (6).
|
||
Assay Type |
Competitive ELISA, Immobilized antigen
|
||
Detection Method |
The Mouse/Rat Testosterone ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA), based on the principle of competitive binding. An unknown amount of testosterone present in the sample and a defined amount of testosterone conjugated to horseradish peroxidase compete for the binding sites of testosterone antiserum coated to the wells of a microplate. After one-hour incubation on a shaker the microplate is washed four times. After addition of the substrate solution the concentration of testosterone is inversely proportional to the optical density measured.
|
||
Reactivity |
Mouse, Rat
|
||
Sample Type |
Serum, plasma
|
||
Sensitivity |
0.066 ng/ml
|
||
Assay Range |
0.1-25 ng / mL
|
||
Kit Components |
1. Microtiterplate, 12 x 8 (break apart) strips with 96 wells; Wells coated with anti-testosterone antibody.
2. Calibrator 0, 1 vial, 0.3 ml, ready to use
3. Calibrator (Calibrator 1-5), 5 vials, 0.3 ml each, ready to use;
Concentrations: 0.1 – 0.4 – 1.5 – 6.0 – 25.0 ng/ml
4. Incubation Buffer, 1 vial 11 ml, ready to use;
5. Enzyme Conjugate, 1 vial, 7 ml, ready to use; Testosterone conjugated to horseradish peroxidise.
6. Substrate Solution, 1 vial, 22 ml, ready to use;
contains tetramethylbenzidine (TMB) and hydrogen peroxide in a buffered matrix.
7. Stop Solution, 1 vial, 7 ml, ready to use; contains 2 N Hydrochloric Acid solution.
8. Wash Solution, 1 vial, 50 ml (10X concentrated).
|
||
Other Reagents Required |
§Centrifuge
§A microtiter plate reader capable for endpoint measurement at 450 nm
§Microplate mixer operating more than 600 rpm
§Vortex mixer
§ Calibrated variable precision micropipettes (10 ul, 50 ul, 100 ul, 200 ul).
§Absorbent paper
§ Distilled or deionized water
§Timer
§Semi logarithmic graph paper or software for data reduction
|
||
Preparation |
All reagents should be at room temperature before use.
Wash Solution:
Dilute 50 ml of 10X concentrated Wash Solution with 450 ml deionized water to a final volume of 500 ml.
The diluted Wash Solution is stable for at least 3 months at room temperature.
1. Disposal of the kits
The disposal of the kit must be made according to the national regulations. Special information for this product is given in the Material Safety Data Sheet.
2. amaged test kits
In case of any severe damage of the test kit or components, BioVendor have to be informed written, latest one week after receiving the kit. Severely damaged single components should not be used for a test run. They have to be stored until a final solution has been found. After this, they should be disposed according to the official regulations.
|
||
Storage & Stability |
When stored at 2°C to 8°C unopened reagents will be stable until expiration date. Do not use reagents beyond this date. Opened reagents must be stored at 2°-8°C. After first opening the reagents are stable for 30 days if used and stored properly.
Microtiter wells must be stored at 2°C to 8°C. Take care that the foil bag is sealed tightly.
Protect TMB-Substrate Solution from light.
|
||
Applications |
The Mouse/Rat Testosterone ELISA is a competitive immunoassay for the measurement of testosterone in rat and mouse serum or plasma. Do not use in diagnostic procedure. For research use only.
|
||
Product Description |
SPECIMEN COLLECTION AND STORAGE INSTRUCTIONS
For determination of rat/mouse Testosterone serum and plasma can be used. The procedure calls for 10 μl matrix per well. The samples should assay immediately or aliquot and stored at -20°C. Avoid repeated freeze-thaw cycles. Samles expected to contain rat/mouse Testosterone concentrations higher than the highest calibrator (25 ng/ml) should be diluted with the zero calibrator before assay. The additional dilution step has to be taken into account for the calculation of the results.
Please note: The use of plasma as specimen can result in a diminished precision of this assay.
ASSAY PROCEDURE
Each run must include a standard curve.
1. Prepare a sufficient number of microplate wells to accommodate calibrators and samples in duplicates.
2. Dispense 10 μl of each Calibrator, Sample and Control with new disposable tips into appropriate wells.
3. Dispense 100 μl of Incubation Buffer into each well.
4. Add 50 μl Enzyme Conjugate into each well.
5. Incubate for 60 minutes at room temperature on a Microplate mixer.
Important Note:
Optimal reaction in this assay is markedly dependent on shaking of the microplate!
6. Discard the content of the wells and rinse the wells 4 times with diluted Wash Solution (300 μl per well). Remove as much Wash Solution as possible by beating the microplate on absorbent paper.
7. Add 200 μl of Substrate Solution to each well.
8. Incubate without shaking for 30 minutes in the dark.
9. Stop the reaction by adding 50 μl of Stop Solution to each well.
10. Determine the absorbance of each well at 450 nm. It is recommended to read the wells within 15 minutes.
CALCULATIONS
1. Calculate the average absorbance values for each set of calibrators, controls and patient samples.
2. Using semi logarithmic graph paper, construct a standard curve by plotting the mean absorbance obtained from each standard against its concentration with absorbance value 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 from the calibration curve.
4. Automated method: The results in the IFU have been calculated automatically using a 4 PL (4 Parameter Logistics) curve fit. 4 Parameter Logistics is the preferred calculation method. Other data reduction functions may give slightly different results.
5. The concentration of the samples can be determined directly from this calibrator curve.
Samples with concentrations higher than that of the highest calibrator have to be further diluted. For the calculation of the concentrations, this dilution factor has to be taken into account.
Conversion to SI units: Testosterone (pg/ml) x 3.47 = pmol/l
|
||
Images |
|
||
Specification |
|||
Quantity |
|
Select | Brand | Catalog No. | Product Name | Pack Size | Type | Field of Research | Specification | Quantity | Price(USD) | |
1 | Leading Biology | EK00487E | Human DCN/Decorin ELISA Kit | 96T | ELISA Kit |
|
$645.00 | Add Ask | ||
2 | Leading Biology | EK00726E | General HS/Heparan sulfate ELISA Kit | 96T | ELISA Kit |
|
$745.00 | Add Ask | ||
3 | Leading Biology | EK00038E | General E2/Estradiol ELISA Kit | 96T | ELISA Kit |
|
$545.00 | Add Ask | ||
4 | Leading Biology | EK0003L | Exendin-4 (Exenatide) ADA | 96T | ELISA Kit |
|
$895.00 | Add Ask | ||
5 | Leading Biology | EK00003E | General Testosterone/Testosterone ELISA Kit | 96T | ELISA Kit |
|
$545.00 | Add Ask | ||
6 | Leading Biology | EK00793E | General GSH/Glutathione ELISA Kit | 96T | ELISA Kit |
|
$745.00 | Add Ask |
Complete this form and click send to ask us a question, request a quote or simply say hello.
You have 0 item in your cart
You have 0 item in your inquiry list