CD33 Antibody, anti-human

Name: CD33 Antibody, anti-human
Availability: InStock

CD33 Antibody, anti-humanExtended Validation

Clone:

AC104.3E3

Type of antibody:

Primary antibodies

Isotype:

mouse IgG1λ

Applications:

FC, MC

Alternative names:

Siglec-3, p67, My9

Flow cytometry applications forCD33 Antibody, anti-human

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 770
VioBlue

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-113-907, 130-113-345

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD33 antibodies as well as with CD14 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

Mass cytometry applications forCD33 Antibody, anti-human

pure

Conjugate:

pure

Products used:

130-108-039

Extended validation for CD33 Antibody, anti-human

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 clones	Overlap in epitope recognition with AC104.3E3
REA775	++
HIM3-4	-
Hu9a (Gemtuzumab Biosimilar)	-
P67.6	+
WM-53	++
Cells were incubated with an excess of purified unconjugated CD33 (AC104.3E3) 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.

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.

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.

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.

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

Comparison of staining pattern on non-fixed and fixed cells using CD33 (AC104.3E3). 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 (AC104.3E3). 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

Overview

Clone AC104.3E3 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.

Alternative names

Siglec-3, p67, My9

Detailed product information

Technical specifications

Clone	AC104.3E3
Clonality	monoclonal
Isotype	mouse IgG1λ
Isotype control	Isotype Control Antibody, mouse IgG1
Host	mouse
Type of antibody	Primary antibodies
Species	human, non-human primate
Cross-reactivity	rhesus monkey ( Macaca mulatta ) , cynomolgus monkey ( Macaca fascicularis )
Antigen	CD33
Alternative names of antigen	Siglec-3, p67, My9
Molecular mass of antigen [kDa]	38
Distribution of antigen	dendritic cells, granulocytes, Langerhans cells, macrophages, mast cells, monocytes, myeloid leukemia cells, NK cells, T cells, basophils, leukemia cells, granulocytes, dendritic cells, Langerhans cells, macrophages, mast cells, monocytes, myeloid leukemia cells, NK cells, T cells, basophils, leukemia cells
Entrez Gene ID	945
RRID	AB_2726123, AB_2726401, AB_2726124, AB_2726397, AB_2726120, AB_2726403, AB_2726126, AB_2726402, AB_2726125, AB_2726398, AB_2726121, AB_2726399, AB_2726122, AB_2660358, AB_2726400

Resources for CD33 Antibody, anti-human

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for CD33 Antibody, anti-human

Publications

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

Applications

Extended validation