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PerCP-Cy™5.5 Mouse Anti-Human CD163
PerCP-Cy™5.5 Mouse Anti-Human CD163

Two-color flow cytometric analysis of CD163 expression on human peripheral blood monocytes.  Whole blood was stained with APC Mouse Anti-Human CD14 antibody (Cat. No. 555399/561708/561383) and either PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control (Cat. No. 550795; dashed line histogram) or PerCP-Cy™5.5 Mouse Anti-Human CD163 antibody (Cat. No. 563887; solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from CD14 positive-gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Two-color flow cytometric analysis of CD163 expression on human peripheral blood monocytes.  Whole blood was stained with APC Mouse Anti-Human CD14 antibody (Cat. No. 555399/561708/561383) and either PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control (Cat. No. 550795; dashed line histogram) or PerCP-Cy™5.5 Mouse Anti-Human CD163 antibody (Cat. No. 563887; solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from CD14 positive-gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Pharmingen™
CD163 antigen; M130; MM130; GHI/61; D11C163A; RM3/1
Human (QC Testing)
Mouse BALB/c IgG1, κ
Glycoprotein preparation from Hairy Cell Leukemia Spleen
Flow cytometry (Routinely Tested)
5 µl
VI M38
9332
AB_2738467
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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

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. 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.
  6. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  7. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Cy is a trademark of GE Healthcare.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563887 Rev. 2
Antibody Details
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GHI/61

The GHI/61 monoclonal antibody specifically binds to human CD163. CD163 is also known as Scavenger receptor cysteine-rich type 1 protein M130 (M130), Hemoglobin scavenger receptor and Macrophage-associated antigen. CD163 is a 110-130 kDa transmembrane glycoprotein. CD163 is a monocyte/macrophage-restricted antigen expressed on the majority of tissue macrophages and peripheral blood monocytes. CD163 belongs to the scavenger receptor superfamily. Its expression on monocytes is upregulated upon cellular activation. CD163 expression reportedly changes on monocytes and macrophages as these cells differentiate. This finding suggests a role for this molecule in the differentiation and/or regulation of monocyte and macrophage function. CD163 may play a role in the clearance and endocytosis of hemoglobin and haptoglobin complexes by macrophages.

It has been reported (Maniecki et al., 2011) that the presence of calcium impacts the binding affinity of clone GHI/61 to CD163. There is a variation in detecting CD163 positive monocytes when the cells are prepared with different anticoagulants, where heparin was observed to have the highest inhibitory effect on clone GHI/61.

563887 Rev. 2
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-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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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
563887 Rev.2
Citations & References
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Development References (5)

  1. Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
  2. Law SK, Micklem KJ, Shaw JM. A new macrophage differentiation antigen which is a member of the scavenger receptor superfamily. Eur J Immunol. 1993; 23(9):2320-2325. (Biology). View Reference
  3. Maniecki MB, Etzerodt A, Moestrup S, Møller J, Graversen J. Comparative assessment of the recognition of domain-specific CD163 monoclonal antibodies in human monocytes explains wide discrepancy in reported levels of cellular surface CD163 expression. Immunobiology. 2011; 216(8):882-890. (Clone-specific: Flow cytometry, Western blot). View Reference
  4. Pulford K, Micklem K, McCarthy S, Cordell J, Jones M, Mason DY. A monocyte/macrophage antigen recognized by the four antibodies GHI/61, Ber-MAC3, Ki-M8 and SM4. Immunology. 1992; 75(4):588-595. (Immunogen: Blocking, Flow cytometry, Immunoaffinity chromatography, Immunohistochemistry, Immunoprecipitation). View Reference
  5. Pulford K, Micklem K, Thomas J, Jones M, Mason DY. A 72-kD B cell-associated surface glycoprotein expressed at high levels in hairy cell leukaemia and plasma cell neoplasms. Clin Exp Immunol. 1991; 85(3):429-435. (Biology). View Reference
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563887 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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.