"P\u011Bknicov\u00E1, Jana" . "Margaryan, Hasmik" . . . "Problem: Characterization of the sperm protein detected by monoclonal antibody (MoAb) Hs-8, which was prepared in our laboratory by immunization of BALB/c mice with human ejaculated sperms. Method of Study: Indirect immunofluorescence, electrophoresis, Western blotting and protein sequencing were used to characterize this protein. For detailed analysis, commercial mouse monoclonal antibody against human sperm-specific glycolytic enzyme (GAPDHS \u2013 ab57062, Abcam, UK) was used. Results: MoAb Hs-8 is a human sperm antibody that cross-reacts with the Hs-8-related protein in spermatozoa of other mammalian species (boar, mouse). In the immunofluorescence test Hs-8 antibody reacted with a protein localised in the arcosomal part of the sperm head and in the principal piece of the sperm flagellum. MoAb Hs-8 labelled a protein of 44 kDa in immunoblot test in the extract of human sperm. Sequence analysis characterized protein Hs-8 as GAPDS (glyceraldehyde 3-phosphate dehydrohenase-spermatogenic). For this reason commercial mouse monoclonal antibody against human sperm-specific glycolytic enzyme (GAPDHS) was applied in other tests. Both antibodies labelled similar proteins in immunofluorescence test (the result was proved by colocalization), in electron microscopy and in immunoblot analysis. Conclusion: GAPDHS is a sperm-specific glycolytic enzyme required for energy production during spermatogenesis and for sperm motility. In our previous paper (JP et al, Theriogeology, 2001) we found that monoclonal antibody Hs-8 blocked the sperm-zona pellucida binding. This protein can be important for secondary sperm/oocyte binding. The role of GAPDS in the sperm head has not yet been elucidated." . . "126817" . . "P(GA523/09/1793), P(GAP503/12/1834), Z(AV0Z50520701)" . . "\u010Capkov\u00E1, Jana" . . "RIV/86652036:_____/12:00380394!RIV13-AV0-86652036" . . . . "Characterization of human sperm protein recognized by monoclonal antibody HS-8" . . "3"^^ . . . . . . "monoclonal antibody; GAPDHS; human sperm proteins; mass spectrometry"@en . "Characterization of human sperm protein recognized by monoclonal antibody HS-8"@en . "Characterization of human sperm protein recognized by monoclonal antibody HS-8" . "3"^^ . "Problem: Characterization of the sperm protein detected by monoclonal antibody (MoAb) Hs-8, which was prepared in our laboratory by immunization of BALB/c mice with human ejaculated sperms. Method of Study: Indirect immunofluorescence, electrophoresis, Western blotting and protein sequencing were used to characterize this protein. For detailed analysis, commercial mouse monoclonal antibody against human sperm-specific glycolytic enzyme (GAPDHS \u2013 ab57062, Abcam, UK) was used. Results: MoAb Hs-8 is a human sperm antibody that cross-reacts with the Hs-8-related protein in spermatozoa of other mammalian species (boar, mouse). In the immunofluorescence test Hs-8 antibody reacted with a protein localised in the arcosomal part of the sperm head and in the principal piece of the sperm flagellum. MoAb Hs-8 labelled a protein of 44 kDa in immunoblot test in the extract of human sperm. Sequence analysis characterized protein Hs-8 as GAPDS (glyceraldehyde 3-phosphate dehydrohenase-spermatogenic). For this reason commercial mouse monoclonal antibody against human sperm-specific glycolytic enzyme (GAPDHS) was applied in other tests. Both antibodies labelled similar proteins in immunofluorescence test (the result was proved by colocalization), in electron microscopy and in immunoblot analysis. Conclusion: GAPDHS is a sperm-specific glycolytic enzyme required for energy production during spermatogenesis and for sperm motility. In our previous paper (JP et al, Theriogeology, 2001) we found that monoclonal antibody Hs-8 blocked the sperm-zona pellucida binding. This protein can be important for secondary sperm/oocyte binding. The role of GAPDS in the sperm head has not yet been elucidated."@en . "RIV/86652036:_____/12:00380394" . . . . . "Characterization of human sperm protein recognized by monoclonal antibody HS-8"@en . . . "[33D4AA1406C9]" . .