The Pig 4MID® KIT is the first ELISA assay that objectively validates semen quality by quantifying a protein biomarker called proAKAP4 in sperm samples (in ejaculate, chilled or frozen semen in extender or in isolated spermatozoa).
| Product Specifications | |
| Research Area | Reproductive Biology |
| Reactive Species | Pig |
| Target | Pig proAKAP4 |
| Tested Applications | ELISA |
| Sample Types | Semen |
| Assay Type | Quantitative Assay |
| Physical Format | 12 breakable Strips of 8-wells |
| Shipping Conditions | Room Temperature |
| Storage Conditions | 4 deg. C |
| Sensitivity | 4 ng/mL |
| Assay Time | 2h to 3h |
| Detection Method | ELISA |
| Assay Range | 0 to 150 ng |
| Materials Required (Not Provided) | - 96-well microplate reader measuring absorbance at 450 nm - Horizontal orbital microplate shaker (300 rpm) - Vortex - Multichannel pipette of 300 μL - Pipettes of 20 μL, 200 μL and 1000 μL - Pipette tips - Polypropylene microtubes of 1.5 mL (for sample preparation) - One polypropylene tube of 15 mL (for R6 dilution) - Plastic reagent reservoirs - One glass bottle (for R2 dilution) - Scissors - Pen - Aluminum foil - Ultrapure or double deionized water |
| Product Contents | R1 - Capture Antibody Coated Microplate of 96-wells (12 x 8-well strips) R2 - 1 Bottle of 10x Washing Buffer Solution (30 mL) R3 - 1 Bottle of 1x Dilution Buffer (30 mL) R4 - 7 Vials of Ready-to-Use Standard Solution (0.7 mL per vial) R5 - 1 Bottle of 1x Pig Spermatozoa Lysis Buffer (30 mL) R6 - 1 Vial of Detection Antibody (0.1 mL) R7 - 1 Bottle of Substrate Solution (11 mL) R8 - 1 Bottle 1x Stop Solution (6 mL) R9 - 1 Vial of Positive Control (0.7 mL) R10 - 2 Adhesive Plate Sealers R11 - User Manual Instructions |
| Regulatory Information | RUO |
| Technical Notes | From one to 88 samples can be run simultaneously or individually (12 breakable strips of 8 wells per 96 well microplate / per kit) |
| Application | Testing Sperm Vitality in samples such as Ejaculates, semen in extenders and Isolated spermatozoa |
| Articles & Publications | Carracedo S, Briand-Amirat L, Dordas-Perpinyà M, Ramos Escuredo Y, Delcombel R, Sergeant N and Dele-hedde M (2022) ProAKAP4 protein marker: towards a functional approach to male fertility. Animal Reproduc-tion Science (in Press). Xu Y, Han Q, Ma C, Wang Y, Zhang P, Li C, Cheng X and Xu H (2021) Comparative proteomics and phosphoproteomics analysis reveal the possible breed difference in Yorkshire and Duroc boar spermatozoa. Front Cell Dev Biol. Vol. 9:652809. Carracedo S, Loyens A, Eddarkaoui S, Serteyn D, Malo C, Skidmore L, Briand-Amirat L, Barbotin AL, Maurage CA, Delehedde M and Sergeant N (2020) The sperm specific proAKAP4 polypeptide exhibited conserved functions, localizations and metabolism among mammals. Animal Reproduction Science. Vol. 220 :106448 Sergeant N, Carracedo S, Blommaert D, Aubry S, Jouy N, Lejeune JP, Franck T, Maurage CA, Serteyn D, Buée L and Delehedde M (2020) Proteolysis of proAKAP4 in semen as a regulatory sensor of sperm quality and functionality. Andrology. Vol. 8 (Suppl. 1):44-45. Sergeant N, Blommaert D, Carracedo S, Franck T, Lejeune JP, Gevaert MH, Aubry S, Maurage CA, Serteyn D and Delehedde M (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. Fang X, Huang LL, Xu J, Ma CQ, Chen ZH, Zhang Z, Liao CH, Zheng SX, Huang P, Xu WM, Li N, Sun L (2019) Proteomics and single-cell RNA analysis of Akap4-knockout mice model confirm indispensable role of Akap4 in spermatogenesis. Nixon B, Bernstein I, Cafe SL, Delehedde M, Sergeant S, Eamens AL, Lord T, Dun MD, De Iuliis GN and Bromfield EG (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 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. Dewulf Q, Briand-Amirat L, Eddarkaoui S, Chambonnet F, Delehedde M and Sergeant N (2019) The effects of freeze-thaw cycles and of storage time on the stability of proAKAP4 polypeptide in raw sperm samples: implications for semen analysis assessment in breeding activities. Journal of Dairy & Veterinary Sciences. Vol. 13(3):1-7 Teijeiro JM and Marini PE (2012) The effect of oviductal deleted in malignant brain tumor 1 over porcine sperm is mediated by a signal transduction pathway that involves pro-AKAP4 phosphorylation. Reproduction. Vol. 143:773-785. |
Pig 4MID Sperm Vitality Assay Kit (ELISA)
- Brand: 4BioDx
- Product Code: 4BioDx-4VDX-18K2
- Availability: Pre-Order
Available Options
Tags: proAKAP4, sperm, semen analysis