User:Robert J. Schiemann/Notebook/Lipsick Lab/2010/04/08

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Spermatogenesis

  • Dissected experimental testes in 1x PBS Solution. Store in eppendorf with 1x PBS until dissections are finished.
    • Two tubes, one for 1:1000 mip40 and another for 1:2500 mip40
    • Fix with 4% Formaldehyde PBS Solution
      • 30 minutes
    • Wash 2 x 5 minutes in 1x PBS-T
    • Incubate 30 minutes with PBS 0.6% Triton.
    • Wash once with 1x PBS-T
    • Block in 2% NGS
      • 1hr at RT or O/N at 4C
    • Incubate with primary antibody in 2% NGS
      • 1:1000 or 1:2500 mip40 B from Rabbit
      • O/N at 4C
  • Continued collecting virgins/males for stable control line.


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