Clone:
REA690
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
CD44s, EMCR III, H-CAM, Pgp-1

Extended validation for CD44 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 REA690
G44-26-
BJ18++
C44Mab-5-
IM7 (h/m)-
515+
MEM-85++
DB105++
IM7.8.1 (h/m)-
REAL259++
Cells were incubated with an excess of purified unconjugated CD44 (REA690) 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 CD44. Human peripheral blood mononuclear cells (PBMCs) were stained with CD44 antibodies and with a suitable counterstaining. As a control, CD44 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 CD44. Human peripheral blood mononuclear cells (PBMCs) were stained with CD44 antibodies and with a suitable counterstaining. As a control, CD44 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 CD44. Human peripheral blood mononuclear cells (PBMCs) were stained with CD44 antibodies and with a suitable counterstaining. As a control, CD44 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 CD44. Human peripheral blood mononuclear cells (PBMCs) were stained with CD44 antibodies and with a suitable counterstaining. As a control, CD44 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 CD44. Human peripheral blood mononuclear cells (PBMCs) were stained with CD44 antibodies and with a suitable counterstaining. As a control, CD44 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 CD44. Human peripheral blood mononuclear cells (PBMCs) were stained with CD44 antibodies and with a suitable counterstaining. As a control, CD44 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 CD44. Human peripheral blood mononuclear cells (PBMCs) were stained with CD44 antibodies and with a suitable counterstaining. As a control, CD44 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 CD44. Human peripheral blood mononuclear cells (PBMCs) were stained with CD44 antibodies and with a suitable counterstaining. As a control, CD44 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 CD44. Human peripheral blood mononuclear cells (PBMCs) were stained with CD44 antibodies and with a suitable counterstaining. As a control, CD44 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 CD44. Human peripheral blood mononuclear cells (PBMCs) were stained with CD44 antibodies and with a suitable counterstaining. As a control, CD44 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 CD44 (REA690). 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 CD44 (REA690). 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 CD44 (REA690). 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 CD44 Antibody, anti-human, REAfinity™

Overview

Clone REA690 recognizes the CD44 antigen. CD44 is a marker for many types of cancer stem cells (CSCs), including breast CSCs that possess higher tumorigenicity and metastatic potential, colorectal, pancreatic, and prostate CSCs. In addition, expression was observed in several cancers as well as on carcinoma cell lines. Here, CD44 plays a role in cancer cell migration and matrix adhesion in response to a cellular microenvironment, thus enhancing cellular aggregation and tumor cell growth. CD44 is also expressed on mesodermal cells, such as hematopoietic, fibroblastic, and glial cells.
Additional information: Clone REA690 displays negligible binding to Fc receptors.

Alternative names

CD44s, EMCR III, H-CAM, Pgp-1

Detailed product information

Technical specifications

CloneREA690
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, non-human primate
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
AntigenCD44
Alternative names of antigenCD44s, EMCR III, H-CAM, Pgp-1
Molecular mass of antigen [kDa]79
Distribution of antigenstem cells, chondrocytes, endothelial cells, epithelial cells, leukocytes, lymphocytes, myeloid leukemia cells, red blood cells, T cells, cancer stem cells, CNS cells, mesenchymal stem cells, plasma cells, ES and iPS cells, bone marrow, kidney, skeletal muscle, skin
Entrez Gene ID960
RRIDAB_2726117, AB_2726395, AB_2726118, AB_2726391, AB_2726114, AB_2726396, AB_2726119, AB_2733231, AB_2733232, AB_2726392, AB_2726115, AB_2726393, AB_2726116, AB_2751813, AB_2751757, AB_2726394

References for CD44 Antibody, anti-human, REAfinity™

Publications

  1. Al-Hajj, M. et al. (2003) Prospective identification of tumorigenic breast cancer cells. Proc. Natl. Acad. Sci. U.S.A. 100: 3983-3988
  2. Dalerba, P. et al. (2007) Phenotypic characterization of human colorectal cancer stem cells. Proc. Natl. Acad. Sci. U.S.A. 104: 10158-10163
  3. Collins, A. T. et al. (2005) Prospective identification of tumorigenic prostate cancer stem cells. Cancer Res. 65: 10946-10951
  4. Aruffo, A. et al. (1990) CD44 is the principal cell surface receptor for hyaluronate. Cell 61: 1303-1313
  5. Patrawala, L. et al. (2006)
    Highly purified CD44
    +
    prostate cancer cells from xenograft human tumors are enriched in tumorigenic and metastatic progenitor cells.
    Oncogene 25: 1696-1708
  6. Wang, L. et al. (2013) Enrichment of prostate cancer stem-like cells from human prostate cancer cell lines by culture in serum-free medium and chemoradiotherapy. Int. J. Biol. Sci. 9(5): 472-479

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