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Alexa Fluor® 647 Mouse Anti-Human CD163
Alexa Fluor® 647 Mouse Anti-Human CD163

Flow cytometric analysis of CD163 expression on human peripheral blood monocytes. Whole blood was stained with either Alexa Fluor® 647 Mouse Anti-Human CD163 antibody (Cat. No. 562669; solid line histogram) or with Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable monocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of CD163 expression on human peripheral blood monocytes. Whole blood was stained with either Alexa Fluor® 647 Mouse Anti-Human CD163 antibody (Cat. No. 562669; solid line histogram) or with Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable 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_2737710
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. 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.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  8. 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.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
562669 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.

562669 Rev. 2
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
562669 Rev.2
Citations & References
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Development References (4)

  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. (Biology). 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). View Reference
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562669 Rev. 2

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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.