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BV421 Rat Anti-Mouse CD68
BV421 Rat Anti-Mouse CD68
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 PE Rat Anti-Mouse F4/80 antibody (Cat. No. 565410) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; dashed line histogram) or BD Horizon BV421 Rat Anti-Mouse CD68 antibody (Cat. No. 566388/566389; 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.
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 PE Rat Anti-Mouse F4/80 antibody (Cat. No. 565410) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; dashed line histogram) or BD Horizon BV421 Rat Anti-Mouse CD68 antibody (Cat. No. 566388/566389; 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
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BD Horizon™
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_2744447
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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  7. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566388 Rev. 1
Antibody Details
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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.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

566388 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
566388 Rev.1
Citations & References
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Development References (6)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
View All (6) View Less
566388 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.