Anti-Mouse CD45R/B220 Monoclonal Antibody (Biotin) for Flow Cytometry - P06800 / P08575

Core Product Specifications and Parameters

Biological Background: CD45R/B220 Function and Localization

• CD45R/B220, encoded by Ptprc, is an isoform of receptor-type tyrosine-protein phosphatase C, also known as leukocyte common antigen (L-CA), lymphocyte antigen 5 (Ly-5), or T200. It is a transmembrane glycoprotein primarily expressed on hematopoietic cells.
• The protein is a key regulator of T-cell activation, acting as a positive co-stimulatory molecule upon binding to DPP4. It contains two tandem intracellular PTPase domains; the first has enzymatic activity, while the second modulates substrate specificity.
• Upon T-cell receptor engagement, CD45 recruits and dephosphorylates signaling molecules such as SKAP1, FYN, and LYN, thereby fine-tuning the activation threshold and preventing autoreactivity.
• CD45 interacts with CLEC10A on antigen-presenting cells at the immunological synapse; this interaction on immature dendritic cells recognizes Tn antigen-carrying CD45 on effector T cells, contributing to peripheral tolerance.
• The B220 isoform is generated by alternative splicing and is predominantly expressed on B lymphocytes and some activated T cells, serving as a classical pan-B cell marker in mice.
• Subcellularly, CD45 localizes to the cell membrane, membrane rafts, and the immunological synapse, where its clustering and exclusion from signaling microdomains influence its activity.
• Post-translational modifications include extensive N- and O-linked glycosylation, which affect its conformation and ligand interactions.
• The mouse protein has a predicted molecular weight of ~144 kDa, but glycosylation and isoform variations result in observed bands between 130 kDa and 144 kDa.

Experimental Guidance and Technical Tips

• For flow cytometry, titrate the biotinylated antibody to determine the optimal concentration for your specific cell population. Include appropriate isotype controls and ensure Fc receptor blocking where necessary.
• Since the antibody is biotinylated, detection requires a fluorophore-conjugated streptavidin or avidin secondary reagent. Choose a streptavidin conjugate with minimal cross-reactivity to your sample species.
• Validate staining specificity on known CD45R/B220-positive and -negative cell lines (e.g., mouse splenocytes or B-cell lines) to confirm performance.
• For multicolor panels, check spectral overlap and use single-stained compensation controls.
• Alternatively, the antibody can be used in other applications after validation, but it is primarily optimized for flow cytometry.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio serves as a comprehensive sourcing partner for IVD raw materials, offering validated antibody pairs, optimized monoclonal and polyclonal antibodies, bulk ancillary reagents, and rare target raw-material sourcing. For researchers focusing on lymphocyte biology and T-cell signaling, our portfolio includes high-quality antibodies like this anti-CD45R/B220 clone, supporting flow cytometry-based immunophenotyping and diagnostic assay development.