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Invitrogen
CD45RA Monoclonal Antibody (HI100), NovaFluor™ Yellow 610, eBioscience™
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CD45RA Antibody (H014T03Y03-A) in Flow
Product Details
H014T03Y03-A
Species Reactivity
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Conjugate
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Product Specific Information
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.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
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 (0.8 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.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 µL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 µL of CellBlox Plus Blocking Buffer per 100 µL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Our internal testing shows that NovaFluor Yellow 610 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.
Excitation: 551 nm; Emission: 614 nm; Laser: 561 nm (Yellow) Laser
Target Information
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.
How to use the Panel Builder
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.
References (0)
Bioinformatics
Protein Aliases: CD45; CD45 antigen; L-CA; Leukocyte common antigen; Ly-5; 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
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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