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Description: The HI100 monoclonal antibody reacts with human CD45RA, a 220 kDa molecule expressed by subpopulations of CD4+ peripheral T lymphocytes, CD8+ peripheral T lymphocytes, and B cells. The CD45RA+ T cell populations are mainly naive/virgin allowing the use of HI100 mAb as a phenotypic marker to discriminate T cell subsets.
Applications Reported: This HI100 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This HI100 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.03 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.
PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 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.
Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
CD45RA is an isoform of the CD45 complex and has restricted expression between different subtypes of lymphoid cells. CD45 (LCA, leukocyte common antigen) is a receptor-type protein tyrosine phosphatase ubiquitously expressed in all nucleated hematopoietic cells, comprising approximately 10% of all surface proteins in lymphocytes. 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 and these isoforms differ in their extracellular domains, however, they share identical transmembrane and cytoplasmic domains. 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. Besides the role in immunoreceptor signaling, CD45 is important in promoting cell survival by modulating integrin-mediated signal transduction pathway and is also involved in DNA fragmentation during apoptosis. CD45 is expressed on cells of the human hematopoietic lineage, with the exception of mature red cells and is not detected on differentiated cells of other tissues. It is likely that CD45 plays an important role in signal transduction, inhibition or upregulation of various immunological functions. Diseases associated with CD45 dysfunction include severe combined immunodeficiency and Hepatitis C.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
Protein Aliases: CD45; CD45 antigen; L-CA; Leukocyte common antigen; Lyt-4; protein tyrosine phosphatase, receptor type, C; protein tyrosine phosphatase, receptor type, c polypeptide; Receptor-type tyrosine-protein phosphatase C; T100; T200; T200 glycoprotein; T200 leukocyte common antigen
Gene Aliases: B220; CD45; CD45R; GP180; L-CA; LCA; LY5; PTPRC; T200
UniProt ID: (Human) P08575
Entrez Gene ID: (Human) 5788
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