Biology Forum › Cell Biology › Biotinylation

last updated by JackBean 14 years, 2 months ago
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    • October 7, 2010 at 7:45 pm #13898
      piglet1
      Participant

      one question about biotinylation. After biotinyled with BIOTIN, and then lysated cells in RIPA buffer and then added strepavidin -beads for 4 hours at 4c, RUN SDS-PAGE gel, probed with the antibody I am interested, should I reprobe the blot with tubulin to proof my experient work or not??
      Thank you

    • October 11, 2010 at 8:57 am #101767
      JackBean
      Participant

      what you mean by reprobe with tubulin?

    • October 12, 2010 at 4:30 pm #101789
      piglet1
      Participant

      Normally we just detect the target proteinafter SDS-PAGE, I mean should we reprobe with anti-tubulin or actin to claim the proteins pulled down with strepavidin beads are only cell memberance fractions.

      thank you

    • October 13, 2010 at 10:30 am #101808
      JackBean
      Participant

      I see. yeah, you could use some controls, by which you should know they will/not be there

    • October 14, 2010 at 7:24 pm #101851
      piglet1
      Participant

      do you know any classic cytoplasmic protein marker beside Tubulin or actin.

      thank you

    • October 21, 2010 at 6:32 am #101955
      JackBean
      Participant

      I would guess some metabolic enzymes, like from glycolysis

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