Background: Human CD34 is a transmembrane glycoprotein which is expressed in
human hematopoietic stem cells (HSCs) and the small- vessel endothelial cells of a
variety of tissues. CD34 plays a critical role as a marker for diagnosis and classification
of leukemia. Anti CD34 antibodies are used for isolation and purification of HSCs from
bone marrow, peripheral blood and cord blood. Objective: To characterize a newly
produced monoclonal antibody against a human CD34 peptide. Methods: Anti CD34
monoclonal antibody (Clone 2C10-D3) was purified from mouse ascitic fluid and
hybridoma cell culture supernatants by affinity chromatography and its immune
reactivity was examined by ELISA. The purified antibody was further characterized
using Western blot and flow cytometry on TF1 (Human Erythroblast) cell line. Results:
ELISA experiment revealed that the antibody recognized CD34 peptide. Western blot
analysis on TF1 cell lysate confirmed the reactivity of the antibody with a 42 KDa
protein. Blocking the antibody with a saturating concentration of specific CD34 peptide
resulted in loss of its activity with TF1 lysate in Western blot. The 2C10-D3 antibody
reacted with TF1 cells in flow cytometry in a similar manner to a commercial anti CD34
monoclonal antibody. Conclusions: Our data suggest that the anti CD34 monoclonal
antibody (Clone 2C10-D3) is an appropriate antibody to study the CD34+ cells by flow
cytometry and Western blot.
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