CD45 Monoclonal Antibody (HI30), PE-Cyanine5, eBioscience™, Invitrogen™

Description

• Description: The HI30 monoclonal antibody reacts with all isoforms of human CD45, also known as Leukocyte Common Antigen (LCA)
• CD45 is expressed by all hematopoietic cells excluding circulating erythrocytes and platelets
• The cytoplasmic portion of CD45 has tyrosine phosphatase enzymatic activity and plays an important role in activation of lymphocytes
• Applications Reported: The HI30 antibody has been reported for use in flow cytometric analysis
• Applications Tested: This HI30 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells
• This can be used at 5 μL (0.015 μg) per test
• A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL
• Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test
• Light sensitivity: This tandem dye is sensitive photo-induced oxidation
• Please protect this vial and stained samples from light
• Fixation: Samples can be stored in IC Fixation Buffer (cat
• 00-8222) (100 μL cell sample + 100 μL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat
• 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation
• Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically
• CD45 (LCA, leukocyte common antigen) is a receptor-type protein tyrosine phosphatase (PTP) ubiquitously expressed in all nucleated hematopoietic cells, comprising approximately 10% of all surface proteins in lymphocytes
• CD45 is absent on non-hematopoietic cell lines, normal and malignant, non-hematopoietic tissues
• CD45 glycoprotein is crucial in lymphocyte development and antigen signaling, serving as an important regulator of Src-family kinases
• CD45 protein exists as multiple isoforms as a result of alternative splicing, differ in their extracellular domains but share identical transmembrane and cytoplasmic domains
• CD45RA is an isoform of the CD45 complex and has restricted expression between different subtypes of lymphoid cells
• CD45 isoforms differ in their ability to translocate into the glycosphingolipid-enriched membrane domains and their expression depends on cell type and physiological state of the cell
• CD45 has been shown to be an essential regulator of T- and B-cell antigen receptor signaling and suppresses JAK kinases to regulate cytokine receptor signaling
• CD45 is also important in promoting cell survival by modulating integrin-mediated signal transduction pathway, DNA fragmentation during apoptosis and inhibition or upregulation of various immunological functions.