title: A novel mAb against a human CD34 peptide reacts with the native protein on CD34+ cells.
abstract: 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.
authors.affiliation: Department of Hematology, Faculty of Allied Medicine, Tehran University of Medical Sciences, Tehran, Iran, e-mail: mahmoodshams87@yahoo.com.
authors.forename: Mahmood
authors.initials: M
authors.lastname: Shams
authors.forename: Mahmood
authors.initials: M
authors.lastname: Jeddi-Tehrani
authors.forename: Farzaneh
authors.initials: F
authors.lastname: Notash Haghighat
authors.forename: Ali Ahmad
authors.initials: AA
authors.lastname: Bayat
authors.forename: Jafar
authors.initials: J
authors.lastname: Mahmoudian
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Rezvani
mesh_terms: D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000378:metabolism*; D018952:Antigens, CD34 / Q000276:immunology; D001853:Bone Marrow / Q000378:metabolism; D002460:Cell Line; D002469:Cell Separation; D042783:Endothelial Cells / Q000378:metabolism*; D000939:Epitopes / Q000276:immunology; D005312:Fetal Blood / Q000378:metabolism; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology; D011485:Protein Binding; D011487:Protein Conformation
chemical_list: D000911:Antibodies, Monoclonal; D018952:Antigens, CD34; D000939:Epitopes; D008565:Membrane Proteins; D010446:Peptide Fragments