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PE-Cy™5 Mouse Anti-Human CD11b
PE-Cy™5 Mouse Anti-Human CD11b
Flow cytometric analysis of CD11b expression on human lymphocytes (Left Plot) or granulocytes (Right Plot). Human whole blood was stained with either PE-Cy™5 Mouse IgG1 κ Isotype Control (Cat. No. 555750; dashed line histogram) or PE-Cy™5 Mouse Anti-Human CD11b (Cat. No. 555389/561686; bold line histogram). Erythrocytes were lysed with Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Fluorescence histograms depicting CD11b (or Ig isotype control) expression were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes or granulocytes. Flow cytometry was performed on a BD FACScan™ system.
Flow cytometric analysis of CD11b expression on human lymphocytes (Left Plot) or granulocytes (Right Plot). Human whole blood was stained with either PE-Cy™5 Mouse IgG1 κ Isotype Control (Cat. No. 555750; dashed line histogram) or PE-Cy™5 Mouse Anti-Human CD11b (Cat. No. 555389/561686; bold line histogram). Erythrocytes were lysed with Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Fluorescence histograms depicting CD11b (or Ig isotype control) expression were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes or granulocytes. Flow cytometry was performed on a BD FACScan™ system.
Product Details
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BD Pharmingen™
MAC-1A; Mac-1; ITGAM; Integrin alpha M; CR3A; CR-3 alpha; Mo1; SLEB6
Human (QC Testing), Rhesus,Cynomolgus,Baboon (Tested in Development)
Mouse IgG1, κ
Human monocytes
Flow cytometry (Routinely Tested)
20 µl
IV M047
AB_395790
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.

Recommended Assay Procedures

  BD™CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. 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™.
  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 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.
  8. 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.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. Cy is a trademark of GE Healthcare.
  11. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  12. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  13. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561686 Rev. 3
Antibody Details
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ICRF44

The ICRF44 monoclonal antibody specifically binds to CD11b, the 165-kDa adhesion glycoprotein that associates with the 95-kDa integrin β2 (CD18) to form the CD11b/CD18 complex, also known as Mac-1 or CR3.  CD11b is a type I transmembrane glycoprotein that is encoded by ITGAM (Integrin alpha M). It is expressed on activated lymphocytes, monocytes, granulocytes, and a subset of NK cells. CD11b functions in cell-cell and cell-substrate interactions and is a receptor for iC3b, CD54 (ICAM-1), CD102 (ICAM-2) and CD50 (ICAM-3). This antibody significantly inhibits polymorphonuclear leukocyte aggregation in response to fMLP.

This clone also cross-reacts with granulocytes, a subset of peripheral blood lymphocytes and some monocytes of baboon, and both rhesus and cynomolgus macaque monkeys. The distribution on lymphocytes and granulocytes is similar to that observed with peripheral blood from normal human donors. There are fewer CD11b-positive monocytes present in the non-human primate blood than in normal human donor samples.

561686 Rev. 3
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
561686 Rev.3
Citations & References
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View product citations for antibody "561686" on CiteAb

Development References (6)

  1. Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
  2. David A, Kacher Y, Specks U, Aviram I. Interaction of proteinase 3 with CD11b/CD18 (beta2 integrin) on the cell membrane of human neutrophils. J Leukoc Biol. 2003; 74(4):551-557. (Biology). View Reference
  3. Hogg N, Horton MA. Myeloid antigens: New and previously defined clusters. In: McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:576-602.
  4. Hogg N, Palmer DG, Revell PA. Mononuclear phagocytes of normal and rheumatoid synovial membrane identified by monoclonal antibodies. Immunology. 1985; 56(4):673-681. (Clone-specific: Immunohistochemistry). View Reference
  5. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  6. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
View All (6) View Less
561686 Rev. 3

 

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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.