User:Vijay/Protocol/western-clfA

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Notes

  • ClfA is a cell wall protein
  • anti-ClfA mAb was gift of Dr. John Vernachio, Inhibitex INC.,
  • Used Normal Rabbit Serum for blocking.
  • Rest, as per our lab protocol.
  • Primary antibody (stock concentration 10ug/ml, 1ul in 10ml, 1:10000, 1ug/ml final concentration)
  • Used regular wash buffer for incubating antibodies.
  • Secondary antibody, Rabbit anti-mouse IgG HRP conjugated from Jackson ImmunoResearch (1:100,000, ie., 1ul of anti-mouse in 100ul of washing buffer and 10ul of that added to 10ml of washing buffer).
  • 5minute exposure is good.