Clone:
REA775
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MC
Alternative names:
Siglec-3, p67, My9

Extended validation for CD33 Antibody, anti-human, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA775
AC104.3E3++
HIM3-4-
Hu9a (Gemtuzumab Biosimilar)-
P67.6+
WM-53++
Cells were incubated with an excess of purified unconjugated CD33 (REA775) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

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 CD33. Human peripheral blood mononuclear cells (PBMCs) were stained with CD33 antibodies and with a suitable counterstaining. As a control, CD33 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 CD33. Human peripheral blood mononuclear cells (PBMCs) were stained with CD33 antibodies and with a suitable counterstaining. As a control, CD33 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 CD33. Human peripheral blood mononuclear cells (PBMCs) were stained with CD33 antibodies and with a suitable counterstaining. As a control, CD33 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 CD33. Human peripheral blood mononuclear cells (PBMCs) were stained with CD33 antibodies and with a suitable counterstaining. As a control, CD33 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 CD33. Human peripheral blood mononuclear cells (PBMCs) were stained with CD33 antibodies and with a suitable counterstaining. As a control, CD33 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 CD33. Human peripheral blood mononuclear cells (PBMCs) were stained with CD33 antibodies and with a suitable counterstaining. As a control, CD33 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 CD33. Human peripheral blood mononuclear cells (PBMCs) were stained with CD33 antibodies and with a suitable counterstaining. As a control, CD33 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 CD33. Human peripheral blood mononuclear cells (PBMCs) were stained with CD33 antibodies and with a suitable counterstaining. As a control, CD33 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
View details
Comparison of staining pattern on non-fixed and fixed cells using CD33 (REA775). 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 CD33 (REA775). 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 CD33 (REA775). 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 CD33 Antibody, anti-human, REAfinity™

Overview

Clone REA775 recognizes the human CD33 antigen, a 67 kDa glycoprotein belonging to the sialoadhesin superfamily. The CD33 antigen is highly expressed on human monocytes but weakly on granulocytes and some – but not all – myeloid dendritic cells. The CD33 antigen is also found on myeloid progenitor cells (CFU-GEMM, CFU GM, CFU-G, BFU-E) but is not expressed on lymphocytes, platelets, erythrocytes, or primitive hematopoietic stem cells.
Additional information: Clone REA775 displays negligible binding to Fc receptors.

Alternative names

Siglec-3, p67, My9

Detailed product information

Technical specifications

CloneREA775
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
Cross-reactivity
cynomolgus monkey (
Macaca fascicularis
)
,
rhesus monkey (
Macaca mulatta
)
AntigenCD33
Alternative names of antigenSiglec-3, p67, My9
Molecular mass of antigen [kDa]38
Distribution of antigenT cells, NK cells, dendritic cells, macrophages, mast cells, Langerhans cells, monocytes, granulocytes, leukemia cells, basophils
Entrez Gene ID945
RRIDAB_2657555, AB_2657556, AB_2657557, AB_2657558, AB_2657559, AB_2657560, AB_2657561, AB_2657562, AB_2657563, AB_2657564, AB_2657565, AB_2657566, AB_2657567, AB_2657568, AB_2657569, AB_2657570, AB_2657571, AB_2657572, AB_2657573, AB_2657574, AB_2657575, AB_2819614, AB_2657554

Resources for CD33 Antibody, anti-human, REAfinity™

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for CD33 Antibody, anti-human, REAfinity™

Publications

  1. Simmons, D. et al. (1988) Isolation of a cDNA encoding CD33, a differentiation antigen of myeloid progenitor cells. J. Immunol. 141(8): 2797-2800
  2. Bernstein, I. D. et al. (1987)
    Treatment of acute myeloid leukemia cells
    in vitro
    with a monoclonal antibody recognizing a myeloid differentiation antigen allows normal progenitor cells to be expressed.
    J. Clin. Invest. 79(4): 1153-1159
  3. Hernández-Caselles, T. et al. (2006) A study of CD33 (SIGLEC-3) antigen expression and function on activated human T and NK cells: two isoforms of CD33 are generated by alternative splicing. J. Leukoc. Biol. 79(1): 46-58

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