A Novel mAb against a Human CD34 Peptide Reacts with the Native Protein on CD34+ Cells

Authors

  • Ali Ahmad Bayat Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
  • Farzaneh Notash Haghighat Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
  • Jafar Mahmoudian Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
  • Mahmood Shams Department of Hematology, Faculty of Allied Medicine, Tehran University of Medical Sciences | Department of Hematology, Faculty of Allied Medicine, Iran University of Medical Sciences
  • Mohammad Reza Rezvani Department of Hematology, Faculty of Allied Medicine, Tehran University of Medical Sciences | Department of Hematology, Faculty of Allied Medicine, Iran University of Medical Sciences | | Immune and Gene Therapy Laboratory, Department of Oncology and Pathology, Karolinska Institutet, Stockholm, Sweden
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.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

a novel mab against a human cd34 peptide reacts with the native protein on cd34+ cells

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: t...

full text

Human CD34+/KDR+ cells are generated from circulating CD34+ cells after immobilization on activated platelets.

OBJECTIVE The presence of kinase-insert domain-containing receptor (KDR) on circulating CD34+ cells is assumed to be indicative for the potential of these cells to support vascular maintenance and repair. However, in bone marrow and in granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood, less than 0.5% of CD34+ cells co-express KDR. Therefore, we studied whether CD34+/KDR+ ...

full text

Human CD34 /KDR Cells Are Generated From Circulating CD34 Cells After Immobilization on Activated Platelets

Objective—The presence of kinase-insert domain-containing receptor (KDR) on circulating CD34 cells is assumed to be indicative for the potential of these cells to support vascular maintenance and repair. However, in bone marrow and in granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood, less than 0.5% of CD34 cells co-express KDR. Therefore, we studied whether CD34 /KDR cel...

full text

Novel transcription factors in human CD34 antigen-positive hematopoietic cells.

Transcription factors (TFs) and the regulatory proteins that control them play key roles in hematopoiesis, controlling basic processes of cell growth and differentiation; disruption of these processes may lead to leukemogenesis. Here we attempt to identify functionally novel and partially characterized TFs/regulatory proteins that are expressed in undifferentiated hematopoietic tissue. We surve...

full text

Detecting human CD34+ and CD34- hematopoietic stem and progenitor cells using a Sysmex automated hematology analyzer.

In clinical medicine, particularly in the newly developing stem cell therapies required to support the practice of regenerative medicine, the measurement of both CD34+ and CD34- hematopoietic stem cells (HSC)/hematopoietic progenitor cells (HPC) is important in obtaining more accurate information about the total HSC/HPC content in various stem/progenitor cell sources. We report the results of a...

full text

Hematopoietic Cells − Cells from Cd34 + Ex Vivo Generation of Cd34

http://bloodjournal.hematologylibrary.org/content/94/12/4053.full.html Updated information and services can be found at: (3094 articles) Hematopoiesis and Stem Cells • Articles on similar topics can be found in the following Blood collections http://bloodjournal.hematologylibrary.org/site/misc/rights.xhtml#repub_requests Information about reproducing this article in parts or in its entirety ma...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 10  issue 4

pages  259- 266

publication date 2013-12-01

By following a journal you will be notified via email when a new issue of this journal is published.

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023