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Determination of the serological sex-specific (Sxs) antigen (“H-Y antigen”) in birds and mammals using high-titer antisera and a sensitive urease ELISA

Summary

A rapid, sensitive, and reproducible enzyme-linked immunosorbent assay (ELISA), for the detection of the serological sex-specific (Sxs) antigen (formerly termed H-Y antigen; see Introduction), is described. This assay uses bovine lestes extract as the solid phase antigen, and high-titer anti-Sxs antisera and a urease-conjugated anti rat-IgG as the first and second antibody respectively. The urea containing substrate causes a pH shift in a positive reaction, which in turn is visualized by the use of bromocresol purple as a pH indicator. The method, and some representative applications of it, are described in detail.

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References

  1. Avrameas S, Ternynck T, Guesdon J-L (1978) Coupling of enzymes to antibodies and antigens. Scand J Immunol [Suppl 7] 8:7–23

  2. Bradley M, Heslop B (1984) An improved urease-ELISA protocol for the screening of monoclonal H-Y antibodies. Proc Univ Otago Med Sch 62:14–16

  3. Bradley M, Heslop BF (1985) Elicitation of a rapid and transient antibody response to H-Y antigen by intrasplenic immunization. Transplantation 39:634–638

  4. Brunner M, Moreira-Filho CA, Wachtel G, Wachtel S (1984) On the secretion of H-Y antigen. Cell 37:615–619

  5. Eichwald EJ, Silmser CR (1955) Untitled communication. Transplant Bull 2:148–149

  6. Fellous M, Günther E, Kemler R, Wiels J, Berger R, Guenet JL, Jakob H, Jacob F (1978) Association of the H-Y male antigen with β2 on human lymphoid and differentiated mouse teratocarcinoma cell lines. J Exp Med 147:58–70

  7. Goldberg EH, Boyse EA, Bennett D, Scheid M, Carswell EA (1971) Serological demonstration of H-Y (male) antigen on mouse sperm. Nature 232:478–480

  8. Goodfellow PN, Andrews PW (1982) Sexual differentiation and H-Y antigen. Nature 295:11–13

  9. Gore-Langton RE, Tung PS, Fritz IB (1983) The absence of specific interactions of Sertoli cell-secreted proteins with antibodies directed against H-Y antigen. Cell 32:289–301

  10. Hall JL, Buskin Y, Wachtel SS (1981) Immunoprecipitation of human H-Y antigen. Hum Genet 58:34–36

  11. Lau YF, Chan K, Kan YW, Goldberg E (1986) Isolation of a malespecific and conserve gene using an anti-H-Y antibody. Am J Hum Genet [Suppl] 39:A142

  12. McLaren A, Simpson E, Tomonari K, Chandler P, Hogg H (1984) Male sexual differentiation in mice lacking H-Y antigen. Nature 312:552–555

  13. Mayerová A, Müller U, Wiberg U, Wolf U, Fraccaro M (1984) Variability in the amount of serologically detectable H-Y antigen. Hum Genet 66:110–112

  14. Moreira-Filho CA, Wachtel SS (1985) Study of H-Y antigen in abnormal sex determination with monoclonal antibody and an ELISA. Am J Med Genet 20:525–534

  15. Müller U, Siebers JW, Zenzes MT, Wolf U (1978) The testis as a secretary organ for H-Y antigen. Hum Genet 45:209–213

  16. Müller U, Zenzes MT, Wolf U, Engel W, Weniger J-P (1979) Appearance of H-W (H-Y) antigen in the gonads of oestradiol sex-reversed male chicken embryos. Nature 280:142–144

  17. Ohno S (1985) The Y-linked testis determining gene and H-Y plasma membrane antigen gene: are they one and the same? Endocr Rev 6:421–431

  18. Scheid M, Boyse EA, Carswell EA, Old LJ (1972) Serologically demonstrable alloantigens of mouse epidermal cells. J Exp Med 135: 938–955

  19. Shalev A, Berezi I, Hamerton JL (1978) Detection and cross-reaction of H-Y antigen by haemagglutination. J Immunogenet 5:303–312

  20. Silvers WK, Gasser DL, Eicher EM (1982) H-Y antigen, serologically detectable male antigen and sex determination. Cell 28:439–440

  21. Simpson E, Chandler P, Hunt R, Hogg H, Tomonari K, McLaren A (1986) H-Y status of X/X Sxr' male mice: in vivo tests. Immunology 57:345–349

  22. Wachtel SS (1983) H-Y antigen and the biology of sex determination. Grune & Stratton, New York

  23. Wachtel SS, Koo GC, Zuckerman EE, Hammerling U, Scheid M, Boyse EA (1974) Serological cross-reactivity between H-Y (male) antigens of mouse and man. Proc Natl Acad Sci USA 71:1215–1218

  24. Wachtel SS, Koo GC, Boyse EA (1975) Evolutionary conservation of H-Y (“male”) antigen. Nature 254:270–272

  25. Waterborg JH, Matthews HR (1984) In: Walker JM (ed). Methods in molecular biology, vol 1: Proteins. Humana Press, Clifton, NJ, pp 1–3

  26. Wiberg UH (1987) Facts and considerations about sex-specific antigens. Hum Genet 76:207–219

  27. Wolf U, Fraccaro M, Mayerová A, Hecht T, Zuffardi O, Hameister H (1980) Turner syndrome patients are H-Y positive. Hum Genet 54:315–318

  28. Zenzes MT, Müller U, Aschmoneit I, Wolf U (1978) Studies on H-Y antigen in different cell fractions of the testis during pubescence. Hum Genet 45:297–303

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Correspondence to Ulf H. Wiberg.

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Bradley, M.P., Ebensperger, C. & Wiberg, U.H. Determination of the serological sex-specific (Sxs) antigen (“H-Y antigen”) in birds and mammals using high-titer antisera and a sensitive urease ELISA. Hum Genet 76, 352–356 (1987). https://doi.org/10.1007/BF00272444

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Keywords

  • Internal Medicine
  • Urea
  • Metabolic Disease
  • Immunosorbent Assay
  • Positive Reaction