PE Mouse Anti-Human CD65

BD Pharmingen™ PE Mouse Anti-Human CD65

Name: PE Mouse Anti-Human CD65
Brand: BD Pharmingen™
SKU: 565858

Clone VIM8 (also known as VIM-8)

(RUO)

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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Product Details

Brand: BD Pharmingen™

Alternative Name: Fucoganglioside Type II: Ceramide dodecasacharide 4c

Reactivity: Human (QC Testing)

Isotype: Mouse IgM, κ

Immunogen: Human THP-1 Cell Line

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl

Workshop Number: IV M23; V MA95

RRID: AB_2739380

Storage Buffer: Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.

Regulatory Status: 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 the dye under optimum conditions and unconjugated antibody and free dye were removed.

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.

Product Notices

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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).
An isotype control should be used at the same concentration as the antibody of interest.
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.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
For U.S. patents that may apply, see bd.com/patents.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

PE Mouse IgM, κ Isotype Control RUO

Size 100 Tests Cat No. 555584

Lysing Solution 10X Concentrate CE_IVD

Size 100 mL Cat No. 349202

Lysing Buffer RUO

Size 100 mL Cat No. 555899

565858 Rev. 2

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. 2

Format Details

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.

Format: PE

Excitation Source: Yellow-Green 488 nm, 532 nm, 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

565858 Rev.2

Citations & References

Development References (7)

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
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. (Clone-specific: Immunofluorescence, Radioimmunoassay). View Reference
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.
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
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
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
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)

565858 Rev. 2

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.