Testosterone ELISA Kit (ab108666)

Testosterone is a biochemical and it should be same in all the species so theoretically this kit would work with testosterone from all the species. We however have tested it with human samples only. Please note the amount of testosterone is higher in urine as compared to serum and plasma samples.

Samples can be diluted in PBS.

This assay will measure total testosterone, free and also protein-bound. To measure just free testosterone, we have another assay, https://www.abcam.com/human-testosterone-free-elisa-kit-ab178663.html.


We recommend running the two controls in duplicate, so those will require 4 wells. The five standards in duplicate will require 10 wells. One more well is needed for the substrate blank.

I can confirm the lab used PBS to dilute serum samples when validating the kit.

This is most likely due to matrix effect which affect measurement of analyte of interest in ELISA when undiluted serum or plasma samples are used. Matrix effect can arise from a number of matrix components such as phospholipids, carbohydrates, and endogenous metabolites (bilirubin), residual formulation components from intraperitoneal (ip), intravenous (iv), or oral dosing (po) experiments; for example, polyethylene glycol (PEG), solutol, polysorbate (Tween 80), etc, an interaction between the analyte of interest and the matrix, such as covalent binding to plasma proteins or the enzymatic degradation of a prodrug.  The easiest way to overcome this is by using diluted samples e.g. 1/2 or 1/10 dilution.

The test measures both bound and free Testosterone.

I confirmed with the lab and unfortunately do not have good news:
The heavily hemolyzed samples would have a great influence on the results. We do not know of any protocol trick to circumvent this and therefore, recommend to collect new samples.
But we would assume that all samples would interfere with the assay at the same level.