Rabbit anti-Human CD34 Monoclonal Antibody for IF/ICC and Flow Cytometry - P28906

Core Product Specifications and Parameters

Biological Background: CD34 Function and Localization

• Hematopoietic progenitor cell antigen CD34, encoded by the CD34 gene, is a well-established marker for hematopoietic stem and progenitor cells and also marks small vessel endothelia.
• Alternative splicing generates multiple isoforms; the canonical isoform has a calculated molecular weight of approximately 41 kDa (UniProt: 40716 Da), while a shorter 35 kDa variant may arise from post-translational processing or isoform differences.
• CD34 is a single-pass type I transmembrane glycoprotein that likely functions as an adhesion molecule, mediating stem cell attachment to the bone marrow extracellular matrix or stromal cells during early hematopoiesis.
• It may serve as a scaffold for lineage-specific glycans, presenting carbohydrate ligands to selectins and facilitating cell–cell interactions.
• Subcellular localization is primarily at the cell membrane, consistent with its role in extracellular adhesion and signaling.
• The protein is subject to extensive post-translational modifications, including N- and O-linked glycosylation (glycoprotein) and phosphorylation, which may regulate its function and binding properties.
• Expression is highly selective: CD34 is found on hematopoietic progenitor cells in bone marrow and peripheral blood, as well as on the endothelium of small vessels in many tissues.
• The extracellular region (including amino acids 170–290 used as immunogen) contains multiple glycosylation sites and is exposed on the cell surface, making it an accessible target for antibody-based detection.

Experimental Guidance and Technical Tips

• The immunogen corresponds to the extracellular region (aa 170–290) of human CD34, ensuring recognition of the native surface antigen; this antibody is validated for IF/ICC and flow cytometry applications.
• For immunofluorescence staining, fix and permeabilize cells appropriately; consider using a non-permeabilizing protocol to detect surface CD34 if desired.
• In flow cytometry, include proper isotype controls and Fc receptor blocking steps to minimize nonspecific binding; titrate the antibody to determine the optimal concentration for your cell system.
• CD34 expression is limited to specific cell types; use well-characterized positive controls (e.g., KG-1a cells, HUVECs, or CD34+ hematopoietic progenitors) and negative controls (e.g., lymphocytes, mature blood cells) to confirm specificity.
• While this conjugate is designed for IF/ICC and FC, if exploring other applications (e.g., IHC, WB), validate the antibody under those conditions and consider potential differences in antigen conformation.

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