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Alexa Fluor® 647 Rat Anti-Human CD115 (CSF-1R)
Alexa Fluor® 647 Rat Anti-Human CD115 (CSF-1R)
Two-parameter flow cytometric analysis of CD115 (CSF-1R) expression on human peripheral blood leucocytes. Whole blood cells were stained with either Alexa Fluor® 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; left panel) or Alexa Fluor® 647 Rat Anti-Human CD115 (CSF-1R) (Cat. No. 564945; 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) characteristics were derived from gated events with the forward and side light-scattering characteristics of viable leucocytes. Flow cytometric analysis was performed on a BD FACSCanto™ II.
Two-parameter flow cytometric analysis of CD115 (CSF-1R) expression on human peripheral blood leucocytes. Whole blood cells were stained with either Alexa Fluor® 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; left panel) or Alexa Fluor® 647 Rat Anti-Human CD115 (CSF-1R) (Cat. No. 564945; 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) characteristics were derived from gated events with the forward and side light-scattering characteristics of viable leucocytes. Flow cytometric analysis was performed on a BD FACSCanto™ II.
Product Details
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BD Pharmingen™
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/test
V MA199
1436
AB_2739022
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.

Recommended Assay Procedures

Please note that immunofluorescent staining of human CD115 is temperature senitive. Staining performed at 4°C gives much stronger immunofluorescent staining intensities upon flow cytometric analysis.

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. 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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564945 Rev. 3
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.

  

564945 Rev. 3
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
564945 Rev.3
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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564945 Rev. 3

Please refer to Support Documents for Quality Certificates


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.