Salivary Testosterone ELISA Kit

Sample

Salivary

Species Reactivity

N/A

Detection Method

cELISA

Intended Use

The Testosterone Enzyme Immunoassay Kit is a competitive immunoassay specifically designed and validated for the quantitative measurement of salivary Testosterone. It is not intended for diagnostic use. It is intended only for research use in humans and some animals.

Contents of Kit

1. Microtitre Plate. Coated with polyclonal anti-Testosterone antibodies. 1 / 96 well
2. Testosterone Standard. 600 pg/mL, in a saliva-like matrix. Serially dilute before use according to Reagent Preparation. Contains: Testosterone, buffer, preservative. 1 vial / 500 μL
3. Testosterone Controls. High / Low. In a saliva-like matrix. Ready to use. Contain: Testosterone, buffer, preservative. 2 vials / 500 μL each
4. Testosterone Enzyme Conjugate Concentrate. Dilute before use with Testosterone Assay Diluent. Contains: Testosterone conjugated to HRP, preservative. 1 vial / 50 μL
5. Testosterone Assay Diluent. Contains: phosphate buffer, pH indicator, preservative. 1 bottle / 30 mL
6. Wash Buffer Concentrate (10×). Dilute before use according to Reagent Preparation. Contains: phosphate buffer, detergent, preservative. 1 bottle / 100 mL
7. TMB Substrate Solution. Non-toxic, ready to use. 1 bottle / 25 mL
8. Stop Solution. 1 bottle / 12.5 mL

Storage

All unopened components of this kit are stable at 2-8°C until the kit' s expiration date.

Precision

The intra-assay precision was determined from the means of 5 samples with 20 replicates of each.

The inter-assay precision was determined from the mean of average duplicates for 5 samples run across two different lots.

Sensitivity

Analytical Sensitivity
The lower limit of sensitivity was determined by interpolating the mean optical density minus 2 SDs of 10 sets of duplicates at the 0 pg/mL level. The minimal concentration of Testosterone that can be distinguished from 0 is 0.458 pg/mL.

Functional Sensitivity
The functional sensitivity is determined as the concentration of saliva samples (tested as 20 replicates and three separate runs) that results in the average lowest concentration with a Coefficient of Variation of 20% or less. The functional sensitivity of the Salivary Testosterone Assay is 0.68 pg/mL.

General Description

estosterone is an anabolic steroid hormone synthesized from androstenedione in the Leydig cells of the testes of males and, in smaller quantities, in the ovaries of females. Small amounts are also secreted by the adrenal glands in both sexes. Testosterone production also occurs in peripheral tissues by conversion of circulating DHEA-S, DHEA, and androstenedione. Testosterone exhibits a diurnal rhythm, with highest levels in the morning and a nadir around midnight.
In men, Testosterone plays an important role in the development of male reproductive tissues including the testes and prostate, as well as promoting secondary sexual characteristics such as increased muscle, bone mass, and hair growth.
In blood, only 1-10% of Testosterone is in its unbound or biologically active form. The remaining Testosterone is bound to serum proteins. Unbound Testosterone enters saliva via intracellular mechanisms, and in saliva the majority of Testosterone is not protein-bound. Salivary Testosterone levels are unaffected by salivary flow rate. The serum-saliva correlation for Testosterone is very high for males, but only modest for females, possibly because women's values often fall near the bottom of the measurable range for both serum and saliva immunoassay kits.

Standard Curve

1. Compute the average optical density (OD) for all duplicate wells.
2. Calculate the percent bound (B/B₀) for each standard, control, and saliva sample by dividing the OD of each well (B) by the average OD for the zero (B₀).
3. Determine the concentrations of the controls and saliva samples by interpolation using data reduction software. We recommend using a 4-parameter non-linear regression curve fit.
4. Samples with Testosterone values greater than 600 pg/mL should be diluted with Testosterone Assay Diluent and rerun for accurate results. If a dilution of the sample is used, multiply the assay results by the dilution factor.
A new Standard Curve must be run with each full or partial plate.