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Alexa Fluor™ 647 Mouse Anti-Human CD65

BD Pharmingen™ Alexa Fluor™ 647 Mouse Anti-Human CD65

Clone VIM8 (also known as VIM-8) (RUO)

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Multiparameter flow cytometric analysis of CD65 expression on Human peripheral blood leukocyte populations.  Human whole blood was stained with either Alexa Fluor™ 647 Mouse IgM, κ Isotype Control (Cat. No. 560806; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human CD65 antibody (Cat. No. 570824/570900). Erythrocytes were lysed with BD FACS Lysing™ Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD65 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.

570824Image1.png

Multiparameter flow cytometric analysis of CD65 expression on Human peripheral blood leukocyte populations.  Human whole blood was stained with either Alexa Fluor™ 647 Mouse IgM, κ Isotype Control (Cat. No. 560806; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human CD65 antibody (Cat. No. 570824/570900). Erythrocytes were lysed with BD FACS Lysing™ Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD65 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.

Product Details
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BD Pharmingen™
Fucoganglioside Type II: Ceramide dodecasacharide 4c
Human (QC Testing)
Mouse IgM, κ
Human THP-1 Cell Line
Flow cytometry (Routinely Tested)
5 µl/test
IV M23; V MA95
AB_3686063
Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® 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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  5. 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).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. An isotype control should be used at the same concentration as the antibody of interest.
  8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  11. For U.S. patents that may apply, see bd.com/patents.
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570900 Rev. 1
Antibody Details
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VIM8

The VIM8 monoclonal antibody specifically binds to the asialylated form of human CD65. CD65 is also known as, Ceramide dodecasacharide 4c. CD65 is expressed on myeloid cells during development. Peripheral blood granulocytes express high levels of CD65when compared with monocytes that express lower levels. CD65 reportedly can serve as a ligand for CD62E (E-Selectin). CD65 has been implicated as a crucial adhesive ligand involved in the extravascular infiltration of acute myeloid leukemic (AML) cells.

570900 Rev. 1
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 670-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.
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Alexa Fluor™ 647
653 nm
669 nm
570900 Rev.1
Citations & References
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View product citations for antibody "570900" on CiteAb

Development References (7)

  1. Easton EW, Schiphorst WE, van Drunen E, van der Schoot CE, van den Eijnden DH. Human myeloid alpha 3-fucosyltransferase is involved in the expression of the sialyl-Lewis(x) determinant, a ligand for E- and P-selectin. Blood. 1993; 81(11):2978-2986. (Clone-specific: Flow cytometry). View Reference
  2. Gooi HC, Hounsell EF, Edwards A, Majdic O, Knapp W, Feizi T. Differences in the fine specificities of monoclonal (Class A) antibodies to human myeloid cells.. Clin Exp Immunol. 1985; 60(1):151-8. (Immunogen: Immunofluorescence, Radioimmunoassay). View Reference
  3. Knapp W., Majdic O, Blanchard D, et al. CDw65 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; 1995:876-882.
  4. Kniep B, Peter-Katalinic J, Müthing J, Majdic O, Pickl WF, Knapp W. The CDw65 monoclonal antibodies VIM-8 and VIM-11 bind to the neutral glycolipid V3FucnLc8Cer.. J Biochem. 1996; 119(3):456-62. (Clone-specific: Flow cytometry, Western blot). View Reference
  5. Noguchi M, Sato N, Sugimori H, Mori K, Oshimi K. A minor E-selectin ligand, CD65, is critical for extravascular infiltration of acute myeloid leukemia cells.. Leuk Res. 2001; 25(10):847-53. (Biology). View Reference
  6. Paietta E, Neuberg D, Bennett JM, et al. Low expression of the myeloid differentiation antigen CD65s, a feature of poorly differentiated AML in older adults: study of 711 patients enrolled in ECOG trials.. Leukemia. 2003; 17(8):1544-50. (Biology). View Reference
  7. Stockinger H. Cluster report: CDw65. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:836-838.
View All (7) View Less
570900 Rev. 1

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.