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BB515 Rat Anti-Human CD115 (CSF-1R)
BB515 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 BB515 Rat IgG1, κ Isotype Control (Cat. No. 564610; left panel) or BB515 Rat Anti-Human CD115 (CSF-1R) (Cat. No. 565346; 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) 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.

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Multiparameter flow cytometric analysis of CD115 (CSF-1R) expression on human peripheral blood leucocytes. Whole blood cells were stained with either BB515 Rat IgG1, κ Isotype Control (Cat. No. 564610; left panel) or BB515 Rat Anti-Human CD115 (CSF-1R) (Cat. No. 565346; 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) 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.

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製品詳細
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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_2739199
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


調製と保管

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™ BB515 under optimum conditions and unconjugated antibody was removed.

推奨アッセイ手順

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

製品通知

  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. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
  6. 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.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565346 Rev. 3
抗体の詳細
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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 BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

565346 Rev. 3
フォーマットの詳細
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BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BB515
Blue 488 nm
490 nm
515 nm
565346 Rev.3
引用&参考文献
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開発者向け参考資料 (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). 参考文献を見る
  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). 参考文献を見る
  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). 参考文献を見る
  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). 参考文献を見る
すべて表示する (5) 表示項目を減らす
565346 Rev. 3

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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.

23-22944-00