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V500 Mouse Anti-Human CD45
V500 Mouse Anti-Human CD45

Flow cytometric analysis of CD45 expression on human lymphocytes. Whole blood was stained with either BD Horizon™ V500 Mouse Anti-Human CD45 (Cat. No. 560777/560779; solid line histogram) or BD Horizon™ V500 Mouse IgG1, κ Isotype Control (Cat. No. 560787; dashed line histgoram). 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 FACSCanto™ flow cytometry system.

Flow cytometric analysis of CD45 expression on human lymphocytes. Whole blood was stained with either BD Horizon™ V500 Mouse Anti-Human CD45 (Cat. No. 560777/560779; solid line histogram) or BD Horizon™ V500 Mouse IgG1, κ Isotype Control (Cat. No. 560787; dashed line histgoram). 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 FACSCanto™ flow cytometry system.

Product Details
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BD Horizon™
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_1937324
Aqueous buffered solution containing protein stabilizer, glycerol 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 with BD Horizon™ V500 under optimum conditions, and unreacted BD Horizon™ V500 was removed.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. BD Horizon V500 has a maximum absorption of 415 nm and maximum emission of 500 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon V500 is covered by one or more of the following US patents: 8,431,416.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560777 Rev. 2
Antibody Details
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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.

The antibody is conjugated to BD Horizon™ V500, which has been developed for use in multicolor flow cytometry experiments and is

available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415 nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors.

When compensating dyes in this spectral range (such as Horizon™ V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls.  Due to spectral differences between cells and beads  in this channel, using BD CompBeads can result in spillover errors for V500 and AmCyan reagents.   Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended.   Different V500 reagents (e.g. CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.

560777 Rev. 2
Format Details
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V500
BD Horizon™ V500 Dye is part of the BD Horizon™ Violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 415-nm and an emission maximum (Em Max) at 499-nm. Driven by BD innovation, V500 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 500 nm (eg, a 525/50 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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V500
Violet 405 nm
415 nm
499 nm
560777 Rev.2
Citations & References
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Development References (8)

  1. 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
  2. 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
  3. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  4. Loken MR, Brosnan JM, Bach BA, Ault KA. Establishing optimal lymphocyte gates for immunophenotyping by flow cytometry. Cytometry. 1990; 11(4):453-459. (Biology). View Reference
  5. 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
  6. Terstappen LW, Levin J. Bone marrow cell differential counts obtained by multidimensional flow cytometry. Blood Cells. 1992; 18(2):311-330. (Biology). View Reference
  7. 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
  8. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
View All (8) View Less
560777 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.