Free Testosterone Saliva ELISA Kit


Human free testosterone saliva ELISA kit is an accurate in vitro quantitative method for detecting unconjugated free testosterone (17beta-hydroxy-4-androstene-3-one) in human saliva samples. This assay has a minimum sensitivity detection limit of 2.2 pg/ml.


Testosterone is a C19 steroid hormone that is produced in the leydig cells of the testes of males and in smaller quantities formed in the ovaries of females. It is known to play vital functions during the development of the male reproductive tissues, bone mass, hair growth, sexual function, supporting increased muscles, stamina, cardiovascular health and the general well-being of the immune system. It is useful in diagnosing and treating disorders that involve androgens. These include primary or secondary hypogonadism, disorders originating from testicular problems, defects of the hypothalamus and pituitary glands. Other disorders are impotence in males, virilization (masculinization) due to tumors, hirsutism (excessive hair) in females, adrenogenital syndroms and polycystic ovaries.

Testosterone displays a diurnal rhythm where the highest levels are seen in the morning and the lowest around midnight. Generally adult males usually have around 10-20 fold higher plasma concentrations present when compared to females. During circulation it is found that the main part is bound to plasma proteins for example sex hormone binding globulin (SHBG) and albumin. Only a small portion approx. 1-2 % is unbound or free, this represents the biologically active form.

So far, every attempts for a direct quantification of free-testosterone using either serum or plasma samples with a commercial immunoassays has failed. The only reliable alternative is using salivary samples to measure it, there have been many publication which have proved that the analysis of free-testosterone using saliva provides clinically valid results even when the concentrations being measured are quite low. Also, using salivary methods it is easier to compensate for the episodic secretion pattern by carrying out multiple sampling over a given period of time. A mixture of these sample can then be used to carry out the analysis procedure, whereas only using single saliva sample per analysis will give arbitrary results, similar to those obtained for serum and plasma samples.


All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • Microtiter Plate (96 Wells): Coated with rabbit polyclonal anti-testosterone antibody.
  • Calibrator 0.
  • Calibrators 1-5: Concentrations 10, 30, 100, 300, 1000 pg/ml.
  • Control 1 (Low) and Control 2 (High).
  • Enzyme Conjugate: Consisting of testosterone conjugated to horseradish peroxidase.
  • Substrate Solution: Tetramethylbenzidine (TMB).
  • Stop Solution: Contains 2 N hydrochloric acid solution.
  • Wash Solution (10x Concentrated).


The minimum sensitivity detection limit of human free testosterone using current testosterone saliva ELISA kit was approximately 2.2 pg/ml. The dynamic range for this assay is 10.0 – 1,000.0 pg/ml.


– Analytical Sensitivity: 2.2 pg/ml
– Specificity (Cross Reactivity): Testosterone (100%), 5α-Dihydrotestosterone (23.3%), Androstenedione (32.2%).
– Assay Dynamic Range: 10 – 1000 pg/ml.
– Intra-Assay Variation: 7.9 – 9.9%
– Inter-Assay Variation: 7.9 – 9.9%
– Recovery: 80.4 – 116.0%
– Linearity: 74 – 117%


  1. Assessing biological maturity: chronological age and the pubertal development scale predict free-testosterone in adolescent males. J Pediatr Endocrinol Metab. (2015) 28 (3-4): 381-6. Hibberd E.E., et al.
  2. Relationships between salivary free testosterone and the expression of force and power in elite athletes. J Sports Med Phys Fitness. (2012) 52 (2): 221-7. Crewther B.T., et al.
  3. A critical evaluation of salivary testosterone as a method for the assessment of serum testosterone. Steroids. (2014) 86: 5-9. Fiers T., et al.


  • Full Name: Free Testosterone Saliva ELISA Kit
  • Reactivity: Human
  • Sample Type: Saliva
  • Sensitivity: 2.2 pg/ml


Online Enquiry Form

"*" indicates required fields

Please check mark information required:


Your secretory IgA ELISA gave good results and I was also really impressed with how quickly we received it.

L. Johnston
PhD Student / University of Glasgow

It is refreshing to know that you have a technical team that is very knowledgeable. I have already recommended your company to other researchers in our department.

Dr. P. Anderson
Lecturer / University College London (UCL)

I am a first time user and found that your instruction manual was very easy to follow. The insulin ELISA assay performed well and I was happy with the results that were generated.

J. Thomas
Senior Technician / Addenbrooke’s Hospital

I carried out a pilot study comparing the performance of many ELISA assay's from different suppliers and found your kits to be one of the better performers. We observed good linearity and tight replicates.

Dr. C. Davies
Lead Scientists / AstraZeneca

You are my first point of contact when I am looking to purchase ELISA. You have such an easy and simple system, yet it is very effective.

A. Shaw
Purchasing / University of Oxford