User:Robert J. Schiemann/Notebook/Lipsick Lab/2010/01/22

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Spermatogenesis

  • With gfp-myb, rfp-h2av samples: Heather performed the following yesterday (1/21/10):
  • Incubate with primary antibody in 2% NGS
    • O/N at 4C
    • Anti-sa with 1:100 anti-sa from guinea pigs
    • Anti-bam with 1:5 anti-bam from mouse
    • Totpro with nothing, continued blocking

Today:

  • Wash 3 x 10 minutes in 1x PBS-T
  • Incubate with secondary antibody in 2% NGS
    • (1:500 of respective secondaries conjugated with 633 fluorophore)
    • Except totpro, remained in 1x PBS-T
    • 2hr at RT
  • Wash 3 x 15 minutes in 1x PBS-T
  • Keep over the weekend in PBS-T.
    • Will stain totpro sample with totpro on Monday and mount the anti-bam and anti-sa samples.