Flow cytometric analysis of CD68 expression by activated F4/80-positive mouse peritoneal macrophages. C57BL/6 mouse thioglycolate-elicited peritoneal exudate cells (PECs) were harvested and fixed and permeabilized using BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV421 Rat Anti-Mouse F4/80 antibody (Cat. No. 565411) and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; dashed line histogram) or PE Rat Anti-Mouse CD68 antibody (Cat. No. 566386/ 566387; solid line histogram) at 0.25 μg/test. The fluorescence histogram showing CD68 expression (or Ig Isotype control staining) was derived from F4/80 positive-gated events with the forward and side light-scatter characteristics of intact PECs. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
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Flow cytometric analysis of CD68 expression by activated F4/80-positive mouse peritoneal macrophages. C57BL/6 mouse thioglycolate-elicited peritoneal exudate cells (PECs) were harvested and fixed and permeabilized using BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV421 Rat Anti-Mouse F4/80 antibody (Cat. No. 565411) and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; dashed line histogram) or PE Rat Anti-Mouse CD68 antibody (Cat. No. 566386/ 566387; solid line histogram) at 0.25 μg/test. The fluorescence histogram showing CD68 expression (or Ig Isotype control staining) was derived from F4/80 positive-gated events with the forward and side light-scatter characteristics of intact PECs. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
Cd68; Macrosialin; Lamp4; gp110; Scard1
Mouse (QC Testing)
Rat IgG2a, κ
Purified Con A Acceptor Glycoproteins from the Mouse P815 Cell Line
Intracellular staining (flow cytometry) (Routinely Tested), Flow cytometry (Tested During Development)
0.2 mg/ml
AB_2744448
Aqueous buffered solution containing ≤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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Product Notices
- An isotype control should be used at the same concentration as the antibody of interest.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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 and Permeabilization Solution RUO
Size
125 mL
Cat No.
554722
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
PE Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
553930
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PE Rat Anti-Mouse CD68 RUO
Size
25 µg
Cat No.
566387
566386 Rev. 1
Antibody Details
FA/11
The FA/11 monoclonal antibody specifically binds to CD68 which is also known as Macrosialin, Lamp4 or Scard1. CD68 is a heavily glycosylated ~ 85-115 kDa type I transmembrane protein that belongs to the lysosomal-associated membrane protein (LAMP) family. Although it is expressed at the cell surface, it is predominantly expressed as an intracellular protein in late endosomes. CD68 is expressed by macrophages, Kupffer cells, histiocytes, microglia, and is considered a useful pan-macrophage marker. It is also expressed by dendritic cells, Langerhans cells and osteoclasts. CD68 expression is upregulated and its glycosylation patterns can be changed by proinflammatory agents. Although CD68 binds to oxidized low-density lipoproteins, its exact biological function is not well defined.
566386 Rev. 1
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is 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 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566386 Rev.1
Citations & References
View product citations for antibody "566386" on CiteAb
Development References (6)
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Ashley JW, Shi Z, Zhao H, Li X, Kesterson RA, Feng X. Genetic ablation of CD68 results in mice with increased bone and dysfunctional osteoclasts.. PLoS ONE. 2011; 6(10):e25838. (Clone-specific: Flow cytometry, Immunofluorescence, Western blot). View Reference
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Kurushima H, Ramprasad M, Kondratenko N, Foster DM, Quehenberger O, Steinberg D. Surface expression and rapid internalization of macrosialin (mouse CD68) on elicited mouse peritoneal macrophages.. J Leukoc Biol. 2000; 67(1):104-8. (Biology). View Reference
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Rabinowitz SS, Gordon S. Macrosialin, a macrophage-restricted membrane sialoprotein differentially glycosylated in response to inflammatory stimuli.. J Exp Med. 1991; 174(4):827-36. (Clone-specific: Depletion, Immunocytochemistry, Immunohistochemistry, Immunoprecipitation, Radioimmunoassay, Western blot). View Reference
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Ramprasad MP, Terpstra V, Kondratenko N, Quehenberger O, Steinberg D. Cell surface expression of mouse macrosialin and human CD68 and their role as macrophage receptors for oxidized low density lipoprotein.. Proc Natl Acad Sci USA. 1996; 93(25):14833-8. (Clone-specific: Flow cytometry). View Reference
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Smith MJ, Koch GL. Differential expression of murine macrophage surface glycoprotein antigens in intracellular membranes.. J Cell Sci. 1987; 87 ( Pt 1):113-9. (Immunogen: Fluorescence microscopy, Immunofluorescence, Immunoprecipitation, Radioimmunoassay). View Reference
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Song L, Lee C, Schindler C. Deletion of the murine scavenger receptor CD68.. J Lipid Res. 2011; 52(8):1542-50. (Clone-specific: Flow cytometry). View Reference
566386 Rev. 1
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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.