Flow cytometric analysis of CD45 expression on human lymphocytes. Whole blood was stained with either Alexa Fluor® 700 Mouse Anti-Human CD45 (Cat. No. 560566; shaded histogram) or Alexa Fluor® 700 Mouse IgG1, κ Isotype Control (Cat. No. 557882; open histogram). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Fluorescent histograms were derived from gated events with the side and forward light-scatter characteristics of viable lymphocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
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Flow cytometric analysis of CD45 expression on human lymphocytes. Whole blood was stained with either Alexa Fluor® 700 Mouse Anti-Human CD45 (Cat. No. 560566; shaded histogram) or Alexa Fluor® 700 Mouse IgG1, κ Isotype Control (Cat. No. 557882; open histogram). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Fluorescent histograms were derived from gated events with the side and forward light-scatter characteristics of viable lymphocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Product Details
BD Pharmingen™
PTPRC; LCA; L-CA; Leukocyte Common Antigen; T200; GP180; LY5
Human (QC Testing)
Mouse IgG1, κ
Human Peripheral Blood Leucocytes
Flow cytometry (Routinely Tested)
5 µl
IV N816
AB_1645452
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
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 to Alexa Fluor® 700 under optimum conditions, and unreacted Alexa Fluor® 700 was removed.
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.
- 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.
- Alexa Fluor® 700 has an adsorption maximum of ~700nm and a peak fluorescence emission of ~720nm. Before staining cells with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Alexa Fluor® 700 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557882
560566 Rev. 2
Antibody Details
HI30
The HI30 monoclonal antibody specifically binds to the 180, 190, 205, 220 kDa protein isoforms of CD45. CD45 is encoded by the PTPRC (Protein tyrosine phosphatase receptor type C) gene. CD45, also known as the leukocyte common antigen (LCA), is a member of the protein tyrosine phosphatase (PTP) family. It is present on all human leukocytes including lymphocytes, monocytes, granulocytes, eosinophils, and thymocytes. CD45 is absent from circulating erythrocytes, platelets, or mature erythroid cells of bone marrow and non-hemopoietic tissues.
560566 Rev. 2
Format Details
Alexa Fluor™ 700
Alexa Fluor™ 700 dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 697 nm and an emission maximum (Em Max) at 719-nm. Alexa Fluor™ 700 is designed to be excited by the Red (627–640-nm) laser and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 700
Red 627-640 nm
697 nm
719 nm
560566 Rev.2
Citations & References
View product citations for antibody "560566" on CiteAb
Development References (13)
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Biffen M, McMichael-Phillips D, Larson T, Venkitaraman A, Alexander D. The CD45 tyrosine phosphatase regulates specific pools of antigen receptor-associated p59fyn and CD4-associated p56lck tyrosine in human T-cells.. EMBO J. 1994; 13(8):1920-1929. (Biology). View Reference
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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. (Biology). View Reference
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Hermiston ML, Xu Z, Weiss A. CD45: a critical regulator of signaling thresholds in immune cells. Annu Rev Immunol. 2003; 21:107-137. (Biology). 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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Loken MR, Brosnan JM, Bach BA, Ault KA. Establishing optimal lymphocyte gates for immunophenotyping by flow cytometry. Cytometry. 1990; 11(4):453-459. (Methodology: Flow cytometry). View Reference
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Phipps DJ, Yousefi S, Branch DR. Increased enzymatic activity of the T-cell antigen receptor-associated fyn protein tyrosine kinase in asymptomatic patients infected with the human immunodeficiency virus. Blood. 1997; 90(9):3603-3612. (Biology). View Reference
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Sieh M, Bolen JB, Weiss A. CD45 specifically modulates binding of Lck to a phosphopeptide encompassing the negative regulatory tyrosine of Lck. EMBO J. 1994; 12(1):315-321. (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
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Terstappen LW, Levin J. Bone marrow cell differential counts obtained by multidimensional flow cytometry. Blood Cells. 1992; 18(2):311-330. (Biology). View Reference
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Trowbridge IS, Thomas ML. CD45: an emerging role as a protein tyrosine phosphatase required for lymphocyte activation and development. Annu Rev Immunol. 1994; 12:85-116. (Biology). View Reference
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Wang Y, Liang L, Esselman WJ. Regulation of the calcium/NF-AT T cell activation pathway by the D2 domain of CD45. J Immunol. 2000; 164(5):2557-2564. (Biology). View Reference
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Wu L, Fu J, Shen SH. SKAP55 coupled with CD45 positively regulates T-cell receptor-mediated gene transcription. Mol Biol Cell. 2002; 22(8):2673-2686. (Biology). View Reference
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
560566 Rev. 2
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.