Multiparameter flow cytometric analysis of CD65 expression on human peripheral blood leucocytes. Human whole blood cells were stained with either PE Mouse IgM Isotype Control (Cat. No. 555584; Left Plot) or PE Mouse Anti-Human CD65 antibody (Cat. No. 565858). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric contour plots showing the correlated expression of CD65 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
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Multiparameter flow cytometric analysis of CD65 expression on human peripheral blood leucocytes. Human whole blood cells were stained with either PE Mouse IgM Isotype Control (Cat. No. 555584; Left Plot) or PE Mouse Anti-Human CD65 antibody (Cat. No. 565858). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric contour plots showing the correlated expression of CD65 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Pharmingen™
Fucoganglioside Type II: Ceramide dodecasacharide 4c
Human (QC Testing)
Mouse IgM, κ
Human Unknown
Flow cytometry (Routinely Tested)
5 µl
IV M23; V MA95
AB_2739380
Aqueous buffered solution containing BSA, protein stabilizer, 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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Product Notices
- 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).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Solution 10X Concentrate CE_IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
PE Mouse IgM, κ Isotype Control RUO
Size
100 Tests
Cat No.
555584
565858 Rev. 1
Antibody Details
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.
565858 Rev. 1
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
565858 Rev.1
Citations & References
Development References (6)
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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
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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.
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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
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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
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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
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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.
565858 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.
