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Horse 4MID® Kit (proAKAP4 marker)

Reference: : 4VDX-18K3

The Horse 4MID® Kit (Ref. 4VDX-18K3) allows to quantify proAKAP4 in horse semen as a biomarker of sperm quality and stallion fertility.

Data sheet




Horse proAKAP4 (all breeds) / Donkey proAKAP4

Analytical range

0 ng to 150 ng

Sample type

Fresh, chilled and frozen semen

More info

Innovative assay to quantify proAKAP4 in stallion semen as a molecular and objective semen quality assessment. 

The Horse 4MID® Kit (Ref. 4VDX-18K3) is the first testing kit that validates objectively semen quality by quantifying a protein named proAKAP4 in samples of horse semen. 

This ELISA kit is the 1st functional testing kit to evaluate sperm quality and male fertility in horse and donkey.

The dosage of the proAKAP4 marker with the Horse 4MID® Kit will bring you: 

  • Qualification of male ejaculate before and after cryopreservation
  • Qualification of each breeding male
  • Best semen for cryopreservation
  • Selection of semen with long lasting motility and functionality at post-thaw
  • Selection of semen with more mature, normal and less-tail abnormalities (indicator of good quality spermatogenesis
  • Selection of best extender for stallion semen preservation

The concentration of the proAKAP4 biomarker is a good indicator of sperm quality and of how the spermatozoa can keep its motility high and efficient overtimes (Sergeant et al., 2019).

proAKAP4 values

High concentrations of proAKAP4 help select semen with long lasting motility and are indicative of good spermatogenesis, giving more mature, more “normal’ spermatozoa and less tail abnormalities ratio in bull semen samples (Bastan and Akcay, 2021).

Technical data

  • Name: Horse 4MID® Kit
  • Reference : 4VDX-18K3
  • This ELISA kit allow to test from 1 to 88 samples simultaneously or individuously (12 breakable strips of 8 well per 96-well microplate / per kit)
  • Working with fresh, chilled or frozen semen in extenders / on isolated spermatozoa
  • Easy to use immunoassay 
  • Easy to interprete / to implement in breeding centers
  • Robust, objective and reproductible method

Research Use Only Product (RUO) / Not for Diagnosis or Therapeutic Use.

Do not hesitate to contact us if you run only one or two samples at the time. We have a special format called the Horse 4MID Kit - BB (ref. 4VDX-18K8BB). Avalaible on request at   

Main references about AKAP4/proAKAP4 marker :

  • Carracedo S, Briand-Amirat L, Dordas-Perpinyà M, Ramos Escuredo Y, Delcombel R, Sergeant N and Delehedde M (2022) ProAKAP4 protein marker: towards a functional approach to male fertility. Animal Reproduction Science. Vol.247(107074):1-20.
  • Smith MZY, Dilger A, Bryan E, Bytnar G, Choi S, Cook K and Kline K (2022) Preliminary Study of Effects of Centrifugation of Stallion Semen on Motility and Pro-AKAP4 Concentration over 72 Hours Post-collection. Journal of Agricultural Science and Technology. Vol (12): 65-72.
  • Dordas-Perpinyà M, Sergeant N, Ruelle I, Bruyas JF, Charreaux C, Michaud S, Carracedo S, Catalán J, Miró J, Delehedde M and Briand-Amirat L (2022) ProAKAP4 semen concentrations as a valuable marker protein of post-thawed semen quality and bull fertility: a retrospective study. Veterinary Sciences. Vol. 9 :224.
  • Dordas-Perpinya M, Yanez I, Catalan J, Eddarkaoui S, Delehedde M, Sergeant N, Bruyas JF, Briand-Amirat L, and Miró J (2022) ProAKAP4 concentrations in stallion and donkey: Comparison with kinetic parameters, motility and concentration. Reproduction in Domestic Animals. Vol. 57(1):61.
  • Dordas-Perpinyà M, Yanez I, Catalan J, Delehedde M, Sergeant N, Bruyas JF, Briand-Amirat L and Miró J (2022) ProAKAP4 correlation with motion parameters and spermatozoa subpopulations in stallion. Reproduction in Domestic Animals. In Press.
  • Dordas-Perpinyà M, Sergeant N, Yanez-Ortiz Y, Catalan J, Delehedde M, Bruyas JF, Briand-Amirat L and Miró J (2022) Characterization of proAKAP4 expression, metabolism and localization in Equus asinus spermatozoa. Reproduction, Fertility and Development. In press.
  • Blommaert D, Sergeant N, Delehedde M, Donnay I, Lejeune JP, Franck T and Serteyn D (2021) First results about ProAKAP4 concentration in stallion semen after cryopreservation in two different freezing media. Cryobiology. Vol. 102:133-135.
  • Bastan I and Akcay E (2021) Quality assessment of frozen bull semen with the precursor A-kinase anchor pro-tein 4 biomarker. Andrologia e14164.
  • Blommaert D, Franck T, Lejeune JF, Selleslagh M, Jouy N, Sergeant N, Serteyn D and Delehedde M (2019) The Horse 4MID® Kit as a promising tool to compare different freezing media to preserve stallion semen. Rev. Bras. Reprod. Anim. Vol 43(2):585.
  • Delehedde M, Carracedo S, Selleslagh M, et al. ProAKAP4 polypeptide as a biomarker of sperm functionality and male fertility disorders. Int J Gynecol and Reprod Sci. (2019);2(1):13-19
  • Sergeant N, Briand-Amirat L, Bencharif D and Delehedde M (2019) The Sperm Specific Protein ProAKAP4 as an innovative marker to evaluate sperm quality and fertility. Journal of Dairy & Veterinary Sciences. Vol. 11:01-19.
  • Nixon B, Bernstein I, Cafe SL, Delehedde M, Sergeant S, Eamens AL, Lord T, Dun MD, De Iuliis GN and Bromfield EG (2019b) A Kinase Anchor Protein 4 is vulnerable to oxidative adduction in male germ cells. Frontiers in Cell and Developmental Biology – Vol. 7:319.
  • Blommaert D, Sergeant N, Delehedde M, Jouy N, Mitchell V, Franck T, Donnay I, Lejeune JP and Serteyn D (2019) Expression, localization, and concentration of A-kinase anchor protein 4 (AKAP4) and its precursor (proAKAP4) in equine semen: Promising marker correlated to the total and progressive motility in thawed spermatozoa. Theriogenology. Vol. 131:52-60.
  • Delehedde M, Blommaert D, Jouy N, Scabello J, Miersman H, Franck T, Serteyn D, Mitchell V and Sergeant S (2018) Concentration of proAKAP4 as a pertinent read-out of sperm quality in mammals. Animal Reproduction Science. Vol. 194:24.
  • Blommaert D, Sergeant N, Delehedde M, Franck T, Lejeune JP and Serteyn D (2018) Significant correlation between the proAKAP4 concentration and the total and progressive motility in stallion sperm after thawing. Journal of Equine Veterinary Science. Vol. 66:43.
  • Sergeant N, Jumeau F, Eddarkaoui S, Sigala J, Dossou GF, Delehedde M, Buee L, Yvoz JF and Mitchell V (2016) Investigating proteomic methods and tools to assess sperm quality. Animal Reproduction Science. Vol. 169:99-135.
  • Turner R, Casas-Dolz P, Schlingmann K and Hameed S (2005) Characterization of an A-kinase anchor protein in equine spermatozoa and examination of the effect of semen cooling and cryopreservation on the binding of that protein to the regulatory subunit of protein kinase-A. American Journal of Veterinary Research. Vol. 66:1055-1056.

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