CD24 Antibody, anti-human, REAfinity™

Name: CD24 Antibody, anti-human, REAfinity™
Availability: InStock

CD24 Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA832

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC, MC

Alternative names:

CD24A, BA-1, HAS

Flow cytometry applications forCD24 Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
Vio Bright R720
VioBlue
VioGreen

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-112-846, 130-112-657

Protocols:

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

Description:

Human peripheral blood cells after erythrocyte lysis were stained with CD24 antibodies or with the corresponding REA Control (S) antibodies (left image) as well as with CD19 antibodies. Flow cytometry was performed 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.

MICS (MACSima Imaging Cyclic Staining) applications forCD24 Antibody, anti-human, REAfinity™

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-112-846, 130-112-657

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forCD24 Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-124-316

Extended validation for CD24 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 clones	Overlap in epitope recognition with REA832
32D12	+
SN3 A5-2H10	+
ML5	+
Cells were incubated with an excess of purified unconjugated CD24 (REA832) 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 CD24. Human peripheral blood mononuclear cells (PBMCs) were stained with CD24antibodies and with a suitable counterstaining. As a control, CD24 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 CD24. Human peripheral blood mononuclear cells (PBMCs) were stained with CD24antibodies and with a suitable counterstaining. As a control, CD24 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 CD24. Human peripheral blood mononuclear cells (PBMCs) were stained with CD24antibodies and with a suitable counterstaining. As a control, CD24 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 CD24. Human peripheral blood mononuclear cells (PBMCs) were stained with CD24antibodies and with a suitable counterstaining. As a control, CD24 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 CD24. Human peripheral blood mononuclear cells (PBMCs) were stained with CD24antibodies and with a suitable counterstaining. As a control, CD24 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 CD24. Human peripheral blood mononuclear cells (PBMCs) were stained with CD24antibodies and with a suitable counterstaining. As a control, CD24 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 CD24 (REA832). 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 CD24 (REA832). 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 CD24 Antibody, anti-human, REAfinity™

Overview

Clone REA832 recognizes the human CD24 antigen, which is also known as heat-stable antigen (HSA). CD24 is a glycosylphosphatidylinositol (GPI)–linked sialoglycoprotein that is anchored to the plasma membrane. CD24 has been identified to be a negative marker for breast cancer stem cells and a positive marker for ovarian or pancreatic cancer stem cells. The CD24 antibody can be used, for example, to differentiate CD44

⁺

CD24

^–

breast cancer stem cells from CD24

⁺

expressing cells from a primary tumor sample. Furthermore, CD24 is found on follicular dendritic cells and different epithelial and hematopoietic cell types.

Additional information: Clone REA832 displays negligible binding to Fc receptors.

Alternative names

CD24A, BA-1, HAS

Detailed product information

Technical specifications

Clone	REA832
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (S), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human
Antigen	CD24
Alternative names of antigen	CD24A, BA-1, HAS
Molecular mass of antigen [kDa]	3
Distribution of antigen	B cells, endothelial cells, epithelial cells, granulocytes, Langerhans cells, monocytes, red blood cells, cancer stem cells, ES and iPS cells, leukemia cells, thymocytes, lung, ovary, pancreas, breast
Entrez Gene ID	100133941
RRID	AB_2656555, AB_2656556, AB_2656557, AB_2656558, AB_2656559, AB_2656560, AB_2656561, AB_2656562, AB_2656563, AB_2656564, AB_2656565, AB_2656566, AB_2656567, AB_2656568, AB_2656569, AB_2656570, AB_2656571, AB_2811647, AB_2904872, AB_2904871, AB_2656554

Resources for CD24 Antibody, anti-human, REAfinity™

Certificates

Please follow this

link

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

References for CD24 Antibody, anti-human, REAfinity™

Publications

Al-Hajj, M. et al. (2003) Prospective identification of tumorigenic breast cancer cells. Proc. Natl. Acad. Sci. U.S.A. 100: 3983-3988
Li, C. P. et al. (2007) Identification of pancreatic cancer stem cells. Cancer Res. 67: 1030-1037
Gao, M. Q. et al. (2010)
CD24
⁺
cells from hierarchically organized ovarian cancer are enriched in cancer stem cells.
Oncogene 29: 2672-2680

Applications

Extended validation