Flow cytometric analysis of CD68 expression by human peripheral blood monocytes. Human peripheral blood mononuclear cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). After washing with BD Perm/Wash™ Buffer (Cat. No. 554723), the cells were stained with either PE-Cy™7 Mouse IgG2b, κ Isotype Control (Cat. No. 560542; dashed line histogram) or PE-Cy™7 Mouse Anti-Human CD68 antibody (Cat. No. 565595; solid line histogram). The fluorescence histogram showing CD68 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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Flow cytometric analysis of CD68 expression by human peripheral blood monocytes. Human peripheral blood mononuclear cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). After washing with BD Perm/Wash™ Buffer (Cat. No. 554723), the cells were stained with either PE-Cy™7 Mouse IgG2b, κ Isotype Control (Cat. No. 560542; dashed line histogram) or PE-Cy™7 Mouse Anti-Human CD68 antibody (Cat. No. 565595; solid line histogram). The fluorescence histogram showing CD68 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
GP110; Macrosialin; SCARD1; Scavenger receptor class D, member 1
Human (QC Testing)
Mouse BALB/c IgG2b, κ
PHA-stimulated Human PBMC
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
IV M149; VI MR23
AB_2739298
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-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed.
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
PE-Cy™7 Mouse IgG2b, κ Isotype Control RUO
Size
0.1 mg
Cat No.
560542
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565595 Rev. 2
Antibody Details
Y1/82A
The Y1/82A monoclonal antibody specifically binds to CD68 which is also known as, Scavenger receptor class D member 1 (SCARD1), Macrosialin, or GP110. CD68 is a cell surface 110 kDa-type I-transmembrane glycoprotein that is primarily expressed in cytoplasmic granules of monocytes, macrophages, dendritic cells, granulocytes, myeloid progenitor cells and, reportedly, a subset of CD34-positive hemopoietic bone marrow progenitor cells. CD68 belongs to the sialomucin family and serves as a scavenger receptor that can bind and internalize oxidized low density lipoproteins (LDL). This antibody is useful in studies of myeloid cell development and function.
565595 Rev. 2
Format Details
PE-Cy7
PE-Cy7 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™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-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.
PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
565595 Rev.2
Citations & References
View product citations for antibody "565595" on CiteAb
Development References (5)
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Davey FR, Cordell JL, Erber WN, Pulford KA, Gatter KC, Mason DY. Monoclonal antibody (Y1/82A) with specificity towards peripheral blood monocytes and tissue macrophages. J Clin Pathol. 1988; 41(7):753-758. (Immunogen: Flow cytometry, Immunohistochemistry). View Reference
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Davey FR, Erber WN, Gatter KC, Mason DY. The use of monoclonal antibody Y1/82A in the identification of acute myeloblastic and monocytic leukemias. Am J Clin Pathol. 1988; 89(1):76-80. (Clone-specific: Immunohistochemistry). View Reference
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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.
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Stockinger H. Cluster report: CD68. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:841-843.
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
565595 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.