Clone:
REA1128
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
VIM-2

Extended validation for CD65 Antibody, anti-human, REAfinity™

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for CD65. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD65 antibodies and with a suitable counterstaining. As a control, CD65 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD65. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD65 antibodies and with a suitable counterstaining. As a control, CD65 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD65. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD65 antibodies and with a suitable counterstaining. As a control, CD65 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD65. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD65 antibodies and with a suitable counterstaining. As a control, CD65 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
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Comparison of staining pattern on non-fixed and fixed cells using CD65 (REA1128). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD65 (REA1128). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using CD65 (REA1128). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD65 Antibody, anti-human, REAfinity™

Overview

Clone REA1128 recognizes the human CD65 antigen, a fucosylated carbohydrate on the cell surface which is also known as ceramide dodecasaccharide or VIM-2. CD65 is formed by fucosylation of the inner lactosamine unit of alpha-2,3-sialylated polylactosamine by alpha-1,3-fucosyltransferase. It is expressed on all myeloid cells during development, highly on granulocytes, and weakly on monocytes in peripheral blood. The sialylated form of CD65 (designated CD65s) appears when the progenitor antigen CD34 disappears, suggesting that this sialylated carbohydrate antigen marks a turning point in normal myeloid differentiation. CD65 is a critical adhesion molecule for extravascular acute myeloid leukemia cell infiltration.
Additional information: Clone REA1128 displays negligible binding to Fc receptors.

Alternative names

VIM-2

Detailed product information

Technical specifications

CloneREA1128
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD65
Alternative names of antigenVIM-2
Distribution of antigengranulocytes, macrophages, monocytes, myeloid cells
RRIDAB_2751716, AB_2751772, AB_2751717, AB_2751771

References for CD65 Antibody, anti-human, REAfinity™

Publications

  1. Lund-Johansen, F. et al. (1992) Activation of human phagocytes through carbohydrate antigens (CD15, sialyl-CD15, CDw17, and CDw65). J. Immunol. 148(10): 3221-3229
  2. Bordessoule, D. et al. (1993) Immunohistological patterns of myeloid antigens: tissue distribution of CD13, CD14, CD16, CD31, CD36, CD65, CD66 and CD67. Br J Haematol 83(3): 370-383
  3. Kniep, B. et al. (1996)
    The CDw65 monoclonal antibodies VIM-8 and VIM-11 bind to the neutral glycolipid V
    3
    FucnLc
    8
    Cer.
    J. Biochem. 119(3): 456-462

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