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BV421 Rat Anti-Human CD115 (CSF-1R)
BV421 Rat Anti-Human CD115 (CSF-1R)

Multiparameter flow cytometric analysis of CD115 (CSF-1R) expression on human peripheral blood leucocytes. Whole blood cells were stained with either BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868; left panel) or BV421 Rat anti-Human CD115 (Cat. No. 565347; right panel). The erythrocytes were lysed with Lysing Buffer (Cat. No. 555899). Two-parameter flow cytometric contour plots showing the correlated expression of CD115 (CSF-1R) [or Ig Isotype control staining] versus Side Light Scatter (SSC-A) signals were derived from gated events with the forward and side light-scattering characteristics of viable leucocytes. Flow cytometric analysis was performed on a BD™ LSR II.

Multiparameter flow cytometric analysis of CD115 (CSF-1R) expression on human peripheral blood leucocytes. Whole blood cells were stained with either BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868; left panel) or BV421 Rat anti-Human CD115 (Cat. No. 565347; right panel). The erythrocytes were lysed with Lysing Buffer (Cat. No. 555899). Two-parameter flow cytometric contour plots showing the correlated expression of CD115 (CSF-1R) [or Ig Isotype control staining] versus Side Light Scatter (SSC-A) signals were derived from gated events with the forward and side light-scattering characteristics of viable leucocytes. Flow cytometric analysis was performed on a BD™ LSR II.

Product Details
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BD Horizon™
CSF-1R; CSF1R; C-FMS; FMS; FIM2; M-CSF-R
Human (QC Testing)
Rat IgG1, κ
Human CD115 Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
V MA199
1436
AB_2739200
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

Note: Please maintain cells around 4°C throughout the staining and analysis procedures for best results.

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 Brilliant Stain Buffer (Cat. No. 563794/566349) or the Brilliant Stain Buffer Plus (Cat. No. 566385).

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. 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.
565347 Rev. 2
Antibody Details
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9-4D2-1E4

The 9-4D2-1E4 monoclonal antibody specifically binds to CD115 which is also known as Colony stimulating factor 1 receptor (CSF-1R) or Macrophage colony-stimulating factor 1 receptor (M-CSFR). This type I transmembrane glycoprotein is a receptor tyrosine kinase (RTK) that belongs to the Ig superfamily. It is expressed on a variety of cells including those committed to the mononuclear phagocyte lineage, such as, monocytes, macrophages, and osteoclasts. CSF-1 binds to and signals through CSF-1R homodimers which undergo tyrosine autophosphorylation and transduce downstream signaling pathways resulting in cytoskeletal reorganization and gene expression. CSF-1R activation stimulates the proliferation, differentiation, and survival of cells within the mononuclear phagocyte system.  Acting through CD115, CSF-1 induces macrophage spreading and motility, and in combination with RANKL, CSF-1 drives the differentiation of mononuclear phagocytes to become osteoclasts. Interleukin-34 (IL-34) is another ligand for CD115 that can induce similar, as well as, some different biological responses by CD115-positive target cells.

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.

565347 Rev. 2
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.
altImg
BV421
Violet 405 nm
407 nm
423 nm
565347 Rev.2
Citations & References
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Development References (5)

  1. Ashmun RA, Look AT, Roberts WM, et al. Monoclonal antibodies to the human CSF-1 receptor (c-fms proto-oncogene product) detect epitopes on normal mononuclear phagocytes and on human myeloid leukemic blast cells. Blood. 1989; 73(3):827-837. (Immunogen: Flow cytometry, Immunoprecipitation). View Reference
  2. Ashmun RA, Look AT, Roussel MF, Sherr CJ. CD115 (CSF-1 receptor) cluster workshop report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:988-989.
  3. Li W, Stanley ER. Role of dimerization and modification of the CSF-1 receptor in its activation and internalization during the CSF-1 response. EMBO J. 1991; 10(2):277-288. (Biology). View Reference
  4. Lin H, Lee E, Hestir K, et al. Discovery of a cytokine and its receptor by functional screening of the extracellular proteome. Science. 2008; 320(5877):807-811. (Biology). View Reference
  5. Sherr CJ, Ashmun RA, Downing JR, et al. Inhibition of colony-stimulating factor-1 activity by monoclonal antibodies to the human CSF-1 receptor. Blood. 1989; 73(7):1786-1793. (Clone-specific: Functional assay). View Reference
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565347 Rev. 2

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