Horse 4MID® Kit (proAKAP4 testing kit)
Product ref. : 4VDX-18K3
The Horse 4MID® Kit (Ref. 4VDX-18K3) allows to quantify proAKAP4 concentrations in horse semen as a marker of sperm quality, long-lasting motility and male fertility.
The Horse 4MID® Kit is the 1st functional testing kit to evaluate sperm quality and male fertility.
Application : in vitro determination of proAKAP4 concentrations in horse sperm (all breeds)
For Research Use Only / Not for Diagnosis or Therapeutic Use
Horse 4MID® Kit Description
The Horse 4MID® Kit (Ref. 4VDX-18K3) is the first test that validates objectively semen quality by quantifying proAKAP4 protein in horse and donkey (all breeds).
The dosage of the proAKAP4 concentrations with the Horse 4MID® Kit brings:
- Objective information on semen quality
- Qualification of each breeding male
- Qualification of each ejaculate
- Qualification of each straw
- Selection of semen with long-lasting motility at post-thaw
- Pertinent parameter to improve sperm cryopreservation
- A functional indicator of male fertility
Application: in vitro quantification of proAKAP4 concentrations in horse and donkey sperm (all breeds).
We have at our catalog since 2022 a BB version of our Horse 4MID® Kit (ref. 4VDX-18K5BB) to analyze from 12 to 24 semen per kit, 1 to 2 samples per strip. Upon request at : contact@4biodx-breeding.com
For Research Use Only / Not for Diagnosis or Therapeutic Use
Reagents provided in the Horse 4MID® Kit
The Horse 4MID® Kit (Ref. 4VDX-18K3) is an ELISA kit that contains all reagents and buffers required for the quantification of proAKAP4 protein in horse / donkey semen in 1 to 88 semen samples (fresh, chilled or frozen semen):
- One Microplate pre-coated with the Capture Antibody
- 7 vials of Standard (ready to use)
- One vial of Positive Control
- One vial of Detection Antibody
- One bottle of the Enzyme Reagent
- One bottle of 10X Wash Buffer
- One bottle of the Sample Diluant
- One bottle of the Acid Stop Solution
- One bottle of Lysis Buffer (specific for horse and donkey semen)
- Two adhesive plate sealers
Do not hesitate to contact us to any technical question or for any quote.
We have at our catalog since 2022 a BB version of our Horse 4MID® Kit (ref. 4VDX-18K3BB) to analyze from 12 to 24 semen per kit, 1 to 2 samples per strip. Upon request at : contact@4biodx-breeding.com
For Research Use Only / Not for Diagnosis or Therapeutic Use
Technical data of the Horse 4MID® Kit
- Name: Horse 4MID® Kit
- Product reference: 4VDX-18K3
- Specificity: Horse and Donkey proAKAP4 (all breeds)
- Method: Quantitative ELISA – 96 well microplate (12 breakable strips of 8 wells)
- Analytical range: 0 ng to 150 ng
- Sample type: Ejaculate / fresh semen, chilled or frozen semen in extenders / isolated spermatozoa
- From one to 88 samples can be run simultaneously or individually (12 breakable strips of 8 wells per microplate / per kit)
Easy to use immunoassay
Easy to interpret: give a number relative to sperm quality and male fertility
Validated by peer-reviewed publications – Available upon request
We have at our catalog since 2022 a BB version of our Horse 4MID® Kit (ref. 4VDX-18K3BB) to analyze from 12 to 24 semen per kit, 1 to 2 samples per strip. Upon request at : contact@4biodx-breeding.com
Research Use Only Product (RUO) / Not for Diagnosis or Therapeutic Use.
Advantages – Horse 4MID® Kit
In horse quantifying proAKAP4 concentrations with the Horse 4MID® Kit is :
- Indicative of the quality of the ejaculate
- Indicative of the sperm motility up to the fertilization site
- Indicative of the quality of frozen straw (quality control)
- Indicative of good spermatogenesis
- Indicative of best semen for cryopreservation
- A tool to select semen with long-lasting motility at post-thaw
- A tool to follow up stallion career
Do not hesitate to contact us for a trial quote or for any questions relating to our innovative approach to assess semen quality and male fertility in stallions !
We have at our catalog since 2022 a BB version of our Horse 4MID® Kit (ref. 4VDX-18K3BB) to analyze from 12 to 24 semen per kit, 1 to 2 samples per strip. Upon request at : contact@4biodx-breeding.com
Overview
The Horse 4MID® Kit as the first molecular test to assess objectively sperm quality and male fertility
The Horse 4MID® Kit detect and quantify the proAKAP4 protein in semen (raw, diluted, fresh, chilled or frozen / isolated spermatoa) from horse and donkey (all breeds).
ProAKAP4 is a marker of semen quality, being a reflect of the good maturation of spermatozoa and good spermatogenesis (Cui et al., 2016; Sergeant et al., 2019). ; Sergeant et al. 2020 ; Delehedde et al. 2019a ; Delehedde et al. 2019b ; Bastan and Akcay, 2021).
ProAKAP4 is the precursor of AKAP4 that is playing a key role in flagellar structure, capacitation and sperm motility (Luconi et al., 2011 ; Delehedde et al. 2019a ; Sergeant et al. 2019 ; Bastan and Akcay, 2021).
Spermatozoa without proAKAP4 / AKAP4 are abnormal, immotile and infertile (Miki et al., 2002 ; Fang et al., 2019).
Oxidative stress decrease proAKAP4 concentration on spermatozoa (Hamada et al., 2013; Nixon et al., 2019 ; Delehedde et al. 2019b).
Sperm from high fertility males have higher expression of proAKAP4 than low fertility males (Peddinti et al., 2008; Ruelle et al., 2019; Sergeant et al., 2019; Griffin et al. 2020).
The concentration of proAKAP4 biomarker measured with the Horse 4MID® Kit is an objective indicator of sperm quality and of how the spermatozoa can keep its motility high and efficient up to fecundation site with fresh semen or at post-thaw !
Examples of applications of the Horse 4MID® Kit in the following references : Blommaert et al. 2021 ; Marta-Dordas et al. 2021 ; Griffin et al. 2020 ; Sergeant et al. 2020 ; Blommaert et al. 2019a ; Blommaert et al. 2019b ; Sergeant et al. 2018).
References
Complete list of publication on the proAKAP4 / AKAP4 system in male fertility upon request. Here below examples of main publications with our products and functional approach on semen quality !
- 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. https://doi.org/10.1016/j.anireprosci.2022.107074
- 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. https://doi.org/10.3390/vetsci9050224
- Kowalczyk A, Gałeska E and Bubel A (2022) The Concentration of ProAKAP4 and Other Indicators of Cryopotential of Spermatozoa Cryopreserved in Extender with Holothuroidea Extract Addition. Animals. Vol. 12:521. https:// doi.org/10.3390/ani12040521
- Dordas-Perpinya et al. (2022) ProAKAP4 concentrations in stallion and donkey: Comparison with kinetic parameters, motility and concentration. Reproduction in Domestic Animals. Vol. 57(1):61.
- Malo C, Carracedo S, Delehedde M, Sergeant N and Skidmore L (2021) Identification of PROAKAP4 in Dromedary sperm and their correlation with monthly semen parameters. Reproduction & Fertility. Vol.2(4):268-279.
- Blommaert et al. (2021) ProAKAP4 concentration in stallion semen after cryopreservation in two different freezing media. Cryobiology. Vol. 102:133-135
- Bastan and Akcay (2021) Quality assessment of frozen bull semen with the precursor A-kinase anchor protein 4 biomarker. Andrologia e14164.
- Sergeant et al (2020) Assessment of the proAKAP4 concentrations as a pertinent sperm parameter to select extenders and preservatives of stallion semen. Animal Reproduction Science. Vol. 220 :106449.
- Carracedo et al. (2020) The sperm specific proAKAP4 polypeptide exhibited conserved functions, localizations and metabolism among mammals. Animal Reproduction Science. Vol. 220 :106448
- Ruelle et al. (2020) ProAKAP4 concentrations in semen as a predictive tool of bull fertility: A preliminary study. Reproduction, Fertility and Development. 32(1):145.
- Griffin et al. (2020) Mass spectrometry reveals distinct proteomic profiles in high- and low-quality stallion spermatozoa. Reproduction. 160(5):695-707.
- Riesco et al. (2020) ProAKAP4 as Novel Molecular Marker of Sperm Quality in Ram: An Integrative Study in Fresh, Cooled and Cryopreserved Sperm. Biomolecules. 10(7):1046.
- Fang et al. (2019) Proteomics and single-cell RNA analysis of Akap4-knockout mice model confirm indispensable role of Akap4 in spermatogenesis. Developmental Biology. 454(2):118-127.
- Blommaert et al. (2019a) 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.
- Blommaert et al. (2019b) 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 et al. (2019) ProAKAP4 polypeptide as a biomarker of sperm functionality and male fertility disorders. Int J Gynecol and Reprod Sci. Vol. 2(1):13-19.
- Delehedde et al (2019b) ProAKAP4 concentrations as an indicator of good spermatogenesis and sperm quality under oxidative stress conditions. Andrology. Vol. 7(1):86.
- Nixon et al. (2019) A Kinase Anchor Protein 4 is vulnerable to oxidative adduction in male germ cells. Frontiers in Cell and Developmental Biology – Vol. 7:319.
- Sergeant et al (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.
- Ugarelli et al. (2018) Identification of six genes related to fertility in alpacas. Animal Reproduction Science. Vol. 194:12-3.
- Luconi et al. (2011) Role of A-kinase anchoring proteins (AKAPs) in reproduction. Frontiers in Bioscience. Vol. 16:1315-1330.
- Hu et al (2009) A-kinase anchoring protein 4 has a conserved role in mammalian spermatogenesis. Reproduction. Vol. 137:645-653.
- Peddinti et al. (2008) Comprehensive proteomic analysis of bovine spermatozoa of varying fertility rates and identification of biomarkers associated with fertility. BMC Systems Biology. Vol. (2)19:1-13.
- Moretti et al. (2007) Localization of AKAP4 and tubulin proteins in sperm with reduced motility. Asian Journal of Andrology. Vol. 9:641-649.
- Turner et al. (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
- Miki et al. (2002) Developmental Biology. Vol. 248: 331-342
Do not hesitate to contact us for a trial quote or for any questions relating to our innovative approach to assess semen quality and male fertility in stallions !
