BD Pharmingen™ PE-Cy7 Mouse Anti-Human CD45
Clone 2D1 (RUO)
Multiparameter flow cytometric analysis of CD45 expression on Human peripheral blood leukocyte populations. Human whole blood was stained with either PE-Cy-7 Mouse IgG1, κ Isotype Control (Cat. No. 565573; Left Plot) or PE-Cy7 Mouse Anti-Human CD45 antibody (Cat. No. 570765/570843; Right Plot) in the presence of BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer (Cat. No. 570002). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD45 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
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Multiparameter flow cytometric analysis of CD45 expression on Human peripheral blood leukocyte populations. Human whole blood was stained with either PE-Cy-7 Mouse IgG1, κ Isotype Control (Cat. No. 565573; Left Plot) or PE-Cy7 Mouse Anti-Human CD45 antibody (Cat. No. 570765/570843; Right Plot) in the presence of BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer (Cat. No. 570002). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD45 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
Product Details
BD Pharmingen™
PTPRC; LCA; L-CA; Leukocyte Common Antigen; T200; GP180; LY5
Human (QC Testing)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl/test
5788
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- 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).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- An isotype control should be used at the same concentration as the antibody of interest.
- PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
MonoBlock™ Leukocyte Staining Buffer RUO
Size
500 Tests
Cat No.
570002
PE-Cy™7 Mouse IgG1 κ Isotype Control RUO
Size
50 µg
Cat No.
565573
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Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
570843 Rev. 1
Antibody Details
2D1
The 2D1 monoclonal antibody recognizes an epitope on all forms of CD45, a tyrosine phosphatase. CD45 is a type I transmembrane glycoprotein that belongs to the Protein Tyrosine Phosphatase (PTP) family. There are several isoforms of CD45 with molecular weights ranging from 180 to 220 kDa. CD45 is expressed on all nucleated cells of hematopoietic origin and is also known as the Leukocyte Common Antigen (LCA). CD45 is most highly expressed on lymphocytes and multiple CD45 isoforms can be coexpressed on individual leucocytes. CD45 plays an important role in regulating leucocyte functions by modifying other signaling molecules.
570843 Rev. 1
Format Details
PE-Cy7
PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-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.
PE-Cy7
Yellow-Green 561 nm
496 nm, 566 nm
781 nm
570843 Rev.1
Citations & References
View product citations for antibody "570843" on CiteAb
Development References (5)
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Bradstock KF, Janossy G, Pizzolo G, et al. Subpopulations of normal and leukemic human thymocytes: an analysis with the use of monoclonal antibodies. J Natl Cancer Inst. 1980; 65(1):33-42. (Immunogen: Immunofluorescence, Immunohistochemistry). View Reference
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Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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Pizzolo G, Sloane J, Beverley P, et al. Differential diagnosis of malignant lymphoma and nonlymphoid tumors using monoclonal anti-leucocyte antibody.. Cancer. 1980; 46(12):2640-7. (Immunogen: Immunofluorescence, Immunohistochemistry). View Reference
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Roederer M, Kantor AB, Parks DR, Herzenberg LA. Cy7PE and Cy7APC: bright new probes for immunofluorescence. Cytometry. 1996; 24(3):191-197. (Biology). View Reference
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Terry LA, Brown MH, Beverley PC. The monoclonal antibody, UCHL1, recognizes a 180,000 MW component of the human leucocyte-common antigen, CD45. Immunology. 1988; 64(2):331-336. (Biology). View Reference
570843 Rev. 1
Please refer to Support Documents for Quality Certificates
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.