Cat 4MID® Kit (proAKAP4 marker testing kit)
Product ref. : 4VDX-19K10
The Cat 4MID® Kit (Ref. 4VDX-19K10) allows to quantify proAKAP4 in samples of feline semen as a biomarker of sperm quality.
The Cat 4MID® Kit is the 1st functional test to evaluate sperm quality and male fertility.
Research Use Only Product (RUO) / Not for Diagnosis or Therapeutic Use.
Cat 4MID® Kit description
The Cat 4MID® Kit (Ref. 4VDX-19K10) is the first immunoassay that validates objectively semen quality by quantifying a protein named proAKAP4 in samples of cat semen.
The dosage of the proAKAP4 biomarker with the Cat 4MID® Kit, will bring :
- Objective information on semen quality
- Qualification of each breeding male
- Qualification of each ejaculate / each straw (control quality)
- Pertinent parameter to improve feline sperm cryopreservation
- Selection of semen with long-lasting motility at post-thaw
- A functional indicator correlated to male fertility
Application : for the in vitro determination of feline proAKAP4 in sperm.
Research Use Only Product (RUO) / Not for Diagnosis or Therapeutic Use.
Reagents provided in the Cat 4MID® Kit
The Cat 4MID® Kit (Ref. 4VDX-19K10) contains all reagents and buffers required for the quantification of proAKAP4 protein in cat semen :
- One Microplate pre-coated with the Capture Antibody
- 7 vials of Standard Solutions (ready to use)
- 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
- Two adhesive plate sealers
Research Use Only Product (RUO) / Not for Diagnosis or Therapeutic Use.
Technical data in the Cat 4MID® Kit
- Name: Cat 4MID® Kit
- Product reference : 4VDX-19K10
- Specificity : Cat proAKAP4
- Method : Sandwich ELISA – 96 well microplate (12 breakable strips of 8 wells)
- Analytical range : 0 ng to 150 ng
- Sample type : Fresh ejaculates, chilled or frozen semen (straws) in extenders / isolated spermatozoa
- From 1 to 88 samples can be run simultaneously or individually (12 breakable strips of 8 wells per plate/per kit)
Easy to use immunoassay
Easy to interpret / give a number relative to sperm quality and male fertility
Research Use Only Product (RUO) / Not for Diagnosis or Therapeutic Use.
Benefits – Cat 4MID® Kit
The Cat 4MID® Kit is:
- Indicative of the semen quality (ejaculate, straws or isolated spermatozoa)
- Indicative of the sperm motility reserve
- Indicative of the quality of frozen straws (control check point)
- A tool to select semen with long-lasting motility at post-thaw
- A support tool for the classification of male breeder for fertility
- A tool to improve cryopreservation of feline semen
Research Use Only Product (RUO) / Not for Diagnosis or Therapeutic Use.
Overview
The Cat 4MID® Kit is the first validated test of sperm quality and male fertility
The Cat 4MID® Kit detect and quantify the proAKAP4 protein in semen (raw, diluted, fresh, chilled or frozen semen / isolated spermatozoa).
ProAKAP4 is the first innovative marker of semen quality, being a reflect of the good maturation of spermatozoa (Cui et al., 2016 ; Delehedde et al. 2019 ; Sergeant et al., 2019) ; 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. 2019).
The precursor proAKAP4 is processed to a “mature” AKAP4 protein that will be active overtimes.
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., 2020).
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; Singh et al., 2019; Fu et al., 2019 ; Griffin et al., 2020).
The concentration of proAKAP4 biomarker measured with the Cat 4MID® Kit is a good indicator of sperm quality and of how the spermatozoa can keep its motility high and efficient overtimes.
References
- 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. In Press.
- 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
- 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.
- 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.
- Bastan and Akcay (2021) Quality assessment of frozen bull semen with the precursor A-kinase anchor protein 4 biomarker. Andrologia e14164.
- 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.
- Carracedo et al. (2020) The sperm specific proAKAP4 polypeptide exhibited conserved functions, localizations and metabolism among mammals. Animal Reproduction Science. Vol. 220 :106448
- Griffin et al. (2020) Mass spectrometry reveals distinct proteomic profiles in high- and low-quality stallion spermatozoa. Reproduction. 160(5):695-707.
- 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.
- Langlade et al. (2020) Assessment of Optimized Frozen/Thawed Semen Samples in Canines with the New A-Kinase Anchor Protein 4 Precursor Biomarker. Biopreserv Biobank. 18(5):409-414.
- Le Couazer et al. (2019a) ProAKAP4 as a valuable marker to assess sperm quality in dogs. Reproduction in Domestic Animals. Vol 54(2):91-92.
- Le Couazer et al. (2019b). Expression of proAKAP4 in dog semen as promising marker of sperm quality. Reproduction in Domestic Animal. Vol 54(S3):73.
- Nixon et al. (2019b) 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.
- Luconi et al. (2011) Role of A-kinase anchoring proteins (AKAPs) in reproduction. Frontiers in Bioscience. Vol. 16:1315-1330.
- Moretti et al. (2007) Localization of AKAP4 and tubulin proteins in sperm with reduced motility. Asian Journal of Andrology. Vol. 9:641-649.
- Miki et al. (2002) Targeted disruption of the AKAP4 gene causes defects in sperm flagellum and motility. Developmental Biology. Vol. 248:331-342.
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