Andrea at Dahl-Chase asked: Is washing with PBA sufficient to ensure that immunoglobulin staining is intrinsic and not extrinsic (cytophilic)? ------------------------------------------ I do not believe that a single wash with PBA (I assume this is PBS with BSA and azide) would be sufficient. Cytophilic Ig is removed either by an acidic wash or by incubating at 37 C for 20-30 min. and then washing with warm media 1-3 times. We incubate ficol/hypaque purified mononuclear cells at 37 C in RPMI with 10% FCS for 30 min and then centrifuge through a cushion of fetal calf serum. On occassion I have just washed F/H cells or lysed whole blood once with PBA and there was still cytophilic Ig present. Tony Bakke, PhD Director, Clinical Immunology and Flow Cytometry Lab Dept of Pathology Oregon Health Sciences University Portland, OR bakkea@ohsu.edu
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:51:42 EST