PerCP-Cy™5.5 Mouse Anti-Human CD73

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BD Pharmingen™ PerCP-Cy™5.5 Mouse Anti-Human CD73

Name: PerCP-Cy™5.5 Mouse Anti-Human CD73
Brand: BD Pharmingen™
SKU: 561260

Clone AD2

(RUO)

Flow cytometric analysis of CD73 expression on human mesenchymal stem cells. Human mesenchymal stem cells (Lonza) at passage 7 were harvested using Accutase™ Cell Detachment Solution (Cat. No. 561527) and stained with PerCP-Cy™5.5 Mouse Anti-Human CD73 antibody (Cat. No. 561260; solid line histogram) or a PerCP-Cy™5.5 mouse IgG1, κ Isotype Control (Cat. No. 550795, dashed line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed using a BD LSRFortessa™ Flow Cytometry System.

Flow cytometric analysis of CD73 on human lymphocytes. Whole blood was stained with PerCP-Cy™5.5 Mouse anti-Human CD73 (Cat. No. 561260; solid line fluorescence histogram) and compared with whole blood stained with FITC Mouse IgG1, κ Isotype Control (Cat. No. 550795; used at a matching concentration; dashed line histogram). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Flow cytometric fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System.

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Product Details

Brand: BD Pharmingen™

Alternative Name: NT5E; 5' nucleotidase; 5'-NT; E5NT; Ecto-5'-nucleotidase; eN; eNT; NT; NT5

Reactivity: Human (QC Testing)

Isotype: Mouse IgG1, κ

Immunogen: Pre-B leukemia cell line

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl

Workshop Number: V B-CD73.3

Entrez Gene ID: 4907

RRID: AB_10896122

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

Product Notices

This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
An isotype control should be used at the same concentration as the antibody of interest.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Accutase™ Cell Detachment Solution RUO

Size 100 mL Cat No. 561527

PerCP-Cy™5.5 Mouse IgG1 κ Isotype Control RUO

Size 0.1 mg Cat No. 550795

561260 Rev. 1

Antibody Details

AD2

The AD2 monoclonal antibody specifically binds to ecto-5'-nucleotidase, a 70 kDa, glycosyl phosphatidylinositol (GPI)-anchored glycoprotein. CD73 is expressed on bone marrow derived multipotent mesenchymal stem cells (MSCs), also sometimes identified as mesenchymal stem cells, and is one of the three positive markers that has been identified by the International Society for Cell Terapy (ISCT) for the minimum criteria for identifying MSCs. Additionally CD73 is expressed on subsets of T and B lymphocytes, follicular dendritic cells, epithelial cells, and endothelial cells. Its expression on lymphocytes increases during T and B cell development. CD73 has enzymatic activity and catalyzes the dephosphorylation of adenosine monophosphate (AMP) converting it to adenosine. It has been suggested that CD73 can mediate costimulatory signals in T cell activation and adhesion of lymphocytes to endothelium.

Due to the expression characteristics of this molecule on different cell types, this PerCP-Cy™5.5 format is only recommended for use on cells expressing high amounts of CD73, such as MSCs.

561260 Rev. 1

Format Details

PerCP-Cy5.5

PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PerCP-Cy5.5

Excitation Source: Blue 488 nm

Excitation Max: 482 nm

Emission Max: 676 nm

561260 Rev.1

Citations & References

Development References (6)

Airas L, Salmi M, Jalkanen S. Lymphocyte-vascular adhesion protein-2 is a novel 70-kDa molecule involved in lymphocyte adhesion to vascular endothelium. J Immunol. 1993; 151(8):4228-4238. (Biology). View Reference
Alam MS, Kurtz CC, Rowlett RM, et al. CD73 is expressed by human regulatory T helper cells and suppresses proinflammatory cytokine production and Helicobacter felis-induced gastritis in mice. J Infect Dis. 2009; 199(4):494-504. (Biology). View Reference
Dominici M, Le Blanc K, Mueller I, et. al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006; 8(4):315-317. (Biology). View Reference
Salazar-Gonzalez JF, Moody DJ, Giorgi JV, Martinez-Maza O, Mitsuyasu RT, Fahey JL. Reduced ecto-5'-nucleotidase activity and enhanced OKT10 and HLA-DR expression on CD8 (T suppressor/cytotoxic) lymphocytes in the acquired immune deficiency syndrome: evidence of CD8 cell immaturity. J Immunol. 1985; 135(3):1778-1785. (Biology). View Reference
Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
Thomson LF, Ruedi JM, Glass A, et al. Production and characterization of monoclonal antibodies to the glycosyl phosphatidylinositol-anchored lymphocyte differentiation antigen ecto-5'-nucleotidase (CD73). Tissue Antigens. 1990; 35(1):9-19. (Biology). View Reference

View All (6)

561260 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.