CD45 Monoclonal Antibody (2D1), Super Bright™ 780, eBioscience™, Invitrogen™

Description

• Description: The 2D1 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: This 2D1 antibody has been reported for use in flow cytometric analysis
• Applications Tested: This 2D1 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells
• This may be used at 5 μL (0.5 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
• Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm
• We recommend using a 780/60 bandpass filter
•  Please make sure that your instrument is capable of detecting this fluorochrome
• In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product No
• 01-2222-42) as compared to single-color stained cells
• In such circumstances, we would recommend setting compensation with cells
• 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.