title: Evaluating in-house anti-serum against B cells with flow cytometry.
abstract: The applications of antibodies, be it monoclonal or polyclonal, in the diagnostic and research fields are well established. The disadvantage is the high cost of commercially available antibodies. In a diagnostic establishment like ours which also functions as a training ground for laboratory related personnel, it is beneficial to be able to produce in-house reagents. Therefore, we have undertaken this project to produce a rabbit polyclonal antibody against B lymphocytes. We found that the rabbit was a good choice because the titre of antibody produced was high and positive reactions were still detected at a dilution of 1:38400. The antibody showed significant positive reaction only with the lymphocyte subpopulation. A positive reaction was observed between the immunized rabbit serum and B lymphocytes but not T lymphocytes. This shows that the antibody was B lymphocyte specific. There was a positive correlation between the percentage of B lymphocytes labelled using the commercial anti-CD19 monoclonal antibody and the in-house polyclonal antibody (n = 13, r = 0.7, p = 0.02). However, the percentage of cells labelled by the in-house polyclonal anti-B was lower than that by the commercial monoclonal anti-CD19. The fluorescence intensity of the polyclonal antibody was lower than that of the monoclonal. In general, the performance of the in-house polyclonal antibody can be considered as satisfactory. The rabbit serum was stored at -20 degrees C and no significant loss of activity was detected for over a period of 19 months.
authors.affiliation: Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur.
authors.forename: S F
authors.initials: SF
authors.lastname: Chin
authors.forename: S K
authors.initials: SK
authors.lastname: Cheong
authors.forename: Y C
authors.initials: YC
authors.lastname: Lim
authors.forename: K L
authors.initials: KL
authors.lastname: Mok
authors.forename: H N
authors.initials: HN
authors.lastname: Hamidah
mesh_terms: D001402:B-Lymphocytes / Q000276:immunology*; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry; D006801:Humans; D007106:Immune Sera
chemical_list: D007106:Immune Sera