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CD73 Monoclonal Antibody (AD2), NovaFluor™ Red 725, eBioscience™
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FIGURE: 1 / 2
CD73 Antibody (H016T03R05-A) in Flow
Normal human peripheral blood cells were unstained (left) or stained with CD73 Monoclonal Antibody, NovaFluor Red 725 (right). All cells were co-stained with CD3 Monoclonal Antibody, eFluor 450 (Product # 48-0038-82). 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
H016T03R05-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG1, kappa
Class
Monoclonal
Type
Antibody
Clone
AD2
Conjugate
Excitation/Emission Max
636/727 nm
View spectra
Form
Liquid
Concentration
0.4 µg/Test
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
Product Specific Information
Description
This AD2 monoclonal antibody reacts with human CD73, a 5'-ectonucleotidase that converts 5'-adenosine monophosphate to adenosine. CD73 is expressed on the surface of endothelial cells, as well as B and T cells, including some CD4+Foxp3+ regulatory T cells. Adenosine production by these cells has been linked to the inhibition of CD4 T cell effector functions such as proliferation and cytokine secretion.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This AD2 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.4 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: 636 nm; Emission: 727 nm; Laser: 633-640 nm (Red) Laser
Target Information
CD73, also known as Ecto-5-prime-nucleotidase or 5-prime-ribonucleotide phosphohydrolase, is an enzyme that catalyzes the conversion of purine 5-prime mononucleotides to nucleosides at neutral pH, with AMP as its preferred substrate. It is composed of a dimer of two identical 70 kDa subunits, externally bound to the plasma membrane via a glycosyl phosphatidyl inositol linkage. CD73 serves as a marker of lymphocyte differentiation and is expressed on a subset of lymphocytes, increasing during lymphocyte maturation. It is found on memory CD4 T cells, which resemble uncommitted primed precursor helper cells (Thpp) capable of differentiating into Th1 or Th2 cells, and is also present on regulatory T cells. CD73 deficiency is associated with various immunodeficiency diseases. Other forms of 5-prime nucleotidase exist in the cytoplasm and lysosomes, distinguishable from CD73 by their substrate affinities, requirement for divalent magnesium ions, activation by ATP, and inhibition by inorganic phosphate. The CD73 gene is localized to chromosome 6q14-q21, and defects in this gene can lead to conditions such as calcification of joints and arteries, and intestinal tuberculosis. Additionally, two transcript variants encoding different isoforms of CD73 have been identified.
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: 5'-deoxynucleotidase; 5'-NT; 5'-nucleotidase; CD73; Ecto-5'-nucleotidase; IMP-specific 5'-nucleotidase; Purine 5-Prime-Nucleotidase; Thymidylate 5'-phosphatase; unnamed protein product
Gene Aliases: CALJA; CD73; E5NT; eN; eNT; NT; NT5; NT5E; NTE
UniProt ID: (Human) P21589
Entrez Gene ID: (Human) 4907
AMP catabolic process
DNA metabolic process
leukocyte cell-cell adhesion
nucleotide catabolic process
response to ATP
ADP catabolic process
ATP metabolic process
adenosine biosynthetic process
negative regulation of inflammatory response
calcium ion homeostasis
inhibition of non-skeletal tissue mineralization
Disclaimer
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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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