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Invitrogen
CD45 Monoclonal Antibody (2D1), NovaFluor™ Violet 800, eBioscience™
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FIGURE: 1 / 2
CD45 Antibody (H005T03V03-A) in Flow
Normal human peripheral blood cells were either left unstained (blue histogram) or stained with CD45 Monoclonal Antibody, NovaFluor Violet 800 (purple histogram). Total viable cells in the lymphocyte gate were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with autofluorescence extraction.
Product Details
H005T03V03-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG1, kappa
Class
Monoclonal
Type
Antibody
Clone
2D1
Conjugate
Excitation/Emission Max
408/800 nm
View spectra
Form
Liquid
Concentration
0.6 µg/Test
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2, with BSA
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
Product Specific Information
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.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
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 (0.6 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 (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, 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
Excitation: 408 nm; Emission: 800 nm; Laser: 405 nm (Violet) Laser
Target Information
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.
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.
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Bioinformatics
Protein Aliases: CD45; CD45 antigen; L-CA; Leukocyte common antigen; protein tyrosine phosphatase, receptor type, C; protein tyrosine phosphatase, receptor type, c polypeptide; Receptor-type tyrosine-protein phosphatase C; T200; T200 glycoprotein; T200 leukocyte common antigen; T220 and B220
Gene Aliases: B220; CD45; CD45R; GP180; L-CA; LCA; LY5; PTPRC; T200
UniProt ID: (Human) P08575
Entrez Gene ID: (Human) 5788
negative regulation of T cell mediated cytotoxicity
negative regulation of cytokine-mediated signaling pathway
hematopoietic progenitor cell differentiation
immunoglobulin biosynthetic process
negative regulation of protein kinase activity
protein dephosphorylation
negative regulation of cell adhesion involved in substrate-bound cell migration
cell surface receptor signaling pathway
dephosphorylation
T cell differentiation
positive regulation of B cell proliferation
peptidyl-tyrosine dephosphorylation
B cell proliferation
positive regulation of T cell proliferation
cell cycle phase transition
positive regulation of protein kinase activity
bone marrow development
stem cell development
T cell receptor signaling pathway
B cell receptor signaling pathway
positive regulation of antigen receptor-mediated signaling pathway
release of sequestered calcium ion into cytosol
defense response to virus
regulation of cell cycle
positive regulation of hematopoietic stem cell migration
positive regulation of stem cell proliferation
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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