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PE-Cy™5 Mouse Anti-Human CD11a
PE-Cy™5 Mouse Anti-Human CD11a

Flow cytometric analysis of CD11a expression on human peripheral blood lymphocytes. Whole blood was stained with either PE-Cy5 Mouse Anti-Human CD11a (Cat. No. 551131; solid line histogram) or PE-Cy5 Mouse IgG1, κ Isotype Control (Cat. No. 550750; dashed line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes.

Flow cytometric analysis of CD11a expression on human peripheral blood lymphocytes. Whole blood was stained with either PE-Cy5 Mouse Anti-Human CD11a (Cat. No. 551131; solid line histogram) or PE-Cy5 Mouse IgG1, κ Isotype Control (Cat. No. 550750; dashed line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes.

Product Details
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BD Pharmingen™
LFA-1α; Lymphocyte (Leukocyte) function-associated antigen 1 α chain; ITGAL
Human (QC Testing), Rhesus, Cynomolgus, Baboon, Dog, Rabbit (Tested in Development)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
20 µl
IV N231
3683
AB_394063
Aqueous buffered solution containing BSA 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 PE-Cy5 (formerly known as BD Cy-Chrome™) under optimum conditions, and unconjugated antibody and free PE-Cy5 were 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. PE-Cy5 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by the 488 nm light of an Argon ion laser and serves as an energy donor, coupled to the cyanine dye Cy5, which acts as an energy acceptor and fluoresces at 670 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in PE-Cy5, thus maximizing its fluorescence emission intensity, minimizing residual emission from PE, and minimizing lot-to-lot variation.
  7. PE-Cy5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the PE-Cy5 tandem fluorochrome, extra care must be taken when using dual-laser cytometers which may directly excite both PE and Cy5™.
  8. PE-Cy5 tandem fluorochromes have been reported to bind some classes of human macrophages and granulocytes via Fc receptors, and PE has been reported to bind to mouse B lymphocytes via Fc receptors. Preincubation of mouse leukocytes with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 can reduce the non-specific binding of PE-Cy5-conjugated reagents to mouse B cells. However, PE-Cy5 conjugated reagents should not be used to stain splenocytes of SJL, NOD, and MRL mice as B lymphocytes and/or other leukocytes have been reported to non-specifically stain regardless of the use of Mouse BD Fc Block™ (the CD72c complex has been implicated for PE-Cy5 binding in these strains). Reagents conjugated to PE, PerCP, PerCP-Cy5.5, APC, and APC-Cy7 tandem fluorochrome can be used on leukocytes from these mouse strains.
  9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  10. Cy is a trademark of GE Healthcare.
  11. 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.
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
551131 Rev. 5
Antibody Details
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HI111

The HI111 monoclonal antibody specifically binds to CD11a, the 180 kDa integrin α chain. This type I transmembrane glycoprotein associates with CD18 (integrin β2) to form the heterodimeric glycoprotein CD11a/CD18. This heterodimer is also known as the lymphocyte (leukocytes) function associated antigen-1 (LFA-1) that is expressed on all leukocytes. LFA-1 is an adhesion molecule involved in lymphocyte and granulocyte functions. LFA-1 mediates adhesion of lymphoid cells to the vascular endothelium in association with its ligand, and the intracellular adhesion molecule-1 (ICAM-1), CD54. Other ligands are ICAM-2 (CD102) and ICAM-3 (CD50).

Clone HI111 also cross reacts with all leukocytes of baboon, and both rhesus and cynomolgus macaque monkeys. The distribution of leukocytes is similar to that observed with peripheral blood leukocytes from normal human donors, with the lymphocyte population also showing a bimodal staining pattern.

551131 Rev. 5
Format Details
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PE-Cy5
PE-Cy5 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™5, with an emission maximum (Em Max) at 670-nm. PE designed to be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 670-nm (e.g., a 670/20-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.
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PE-Cy5
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
670 nm
551131 Rev.5
Citations & References
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Development References (5)

  1. Dustin ML, Springer TA. Lymphocyte function-associated antigen-1 (LFA-1) interaction with intercellular adhesion molecule-1 (ICAM-1) is one of at least three mechanisms for lymphocyte adhesion to cultured endothelial cells. J Cell Biol. 1988; 107(1):321-331. (Biology). View Reference
  2. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  3. Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
  4. Rothlein R, Dustin ML, Marlin SD, Springer TA. A human intercellular adhesion molecule (ICAM-1) distinct from LFA-1. J Immunol. 1986; 137(4):1270-1274. (Biology). View Reference
  5. van Vugt MJ, van den Herik-Oudijk IE, van de Winkle JG. Binding of PE-CY5 conjugates to the human high-affinity receptor for IgG (CD64). Blood. 1996; 88(6):2358-2361. (Biology). View Reference
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551131 Rev. 5

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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.