RE: c-flip antibody

Michelle,
  Check this reference showing nice results of c-flip intracellular staining of CD14+
cells for Flow. Arthritis Rheum 2001; 44: 21-30. Unfortunately the antibody used did not
seem to be a commercial one at time of publication. 
  For intracellular FLIP staining, CD14-labeled MNC were fixed in
  4% neutral buffered formalin, permeabilized with 0.1% Non-
  idet P-40 (NP-40), blocked overnight at 4°C in 90% goat
  serum, and incubated at 4°C for 3–4 hours with rabbit anti-
  FLIP antibody (gift from Ken Walsh [11,14,15]) or control
  rabbit IgG (Sigma). Cells were then incubated with FITClabeled
  goat anti-rabbit antibody (Jackson ImmunoResearch)
  at 4°C for 1–2 hours. FLIP expression was determined in the
  CD14-positive MNC by flow cytometry, and intracellular FLIP
  was quantified by the mean fluorescence intensity.
  Regards,

"Dorn, Michelle E." <MDORN@PARTNERS.ORG> escribió:
Hello All,

I was just wondering if anyone out there has ever looked at the expression of
c-flip (cellular FLICE inhibitory protein) using flow cytometry. Any stories of
success or failures would be greatly appreciated!

Thank you,

Michelle Dorn

Massachussetts General Hospital
Clinical Flow Cytometry Lab
Boston, MA 02114






Guillermo Palao, MD. Ph.D.
Laboratorio de Reumatología
Centro de Investigación
Hospital 12 de Octubre
Avda de Córdoba s/n
Madrid 28041
Spain
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