Clone:
REAL259
Type of antibody:
Releasable antibodies, Primary antibodies, Recombinant antibodies
Applications:
FC, MICS, IHC, IF
Alternative names:
CD44s, EMCR III, H-CAM, Pgp-1

Extended validation for CD44 Antibody, anti-human,
REAlease
®

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 REAL259
G44-26-
BJ18++
C44Mab-5-
IM7 (h/m)-
515+
MEM-85++
DB105++
IM7.8.1 (h/m)-
REA690++
Cells were incubated with an excess of purified unconjugated CD44 (REAL259) 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

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.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using CD44 (REAL259). 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 (REAL259). 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 (REAL259). 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,
REAlease
®

Overview

Clone REAL259 is an antibody fragment derived from the full CD44 antibody molecule. It displays no binding to Fc receptors. The recombinantly engineered antibody fragments are multimerized to form the REAlease Complex to bind markers with high avidity.
Clone REAL259 recognizes the CD44 antigen. CD44 is a marker for many types of cancer stem cells (CSC), 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.
The REAlease Kits consist of the respective fluorochrome-conjugated REAlease Complexes and the REAlease Support Kit for removal of the REAlease Complexes and optional relabeling with different fluorochrome-conjugated REAlease Complexes.

Alternative names

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

Detailed product information

Technical specifications

CloneREAL259
Clonalitymonoclonal
Isotype controlControl Antibody
Hostcell line
Type of antibodyReleasable antibodies, Primary antibodies, Recombinant antibodies
Specieshuman
AntigenCD44
Alternative names of antigenCD44s, EMCR III, H-CAM, Pgp-1
Distribution of antigenfibroblasts, tumor cells, cancer stem cells, tumor
RRIDAB_2819403, AB_2819400, AB_2751287, AB_2751279, AB_2784193, AB_2801703, AB_2811698, AB_2801702

References for CD44 Antibody, anti-human,
REAlease
®

Publications

  1. Aruffo, A. et al. (1990) CD44 is the principal cell surface receptor for hyaluronate. Cell 61: 1303-1313
  2. Al-Hajj, M. et al. (2003) Prospective identification of tumorigenic breast cancer cells. Proc. Natl. Acad. Sci. U.S.A. 100: 3983-3988
  3. Collins, A. T. et al. (2005) Prospective identification of tumorigenic prostate cancer stem cells. Cancer Res. 65: 10946-10951
  4. 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
  5. Dalerba, P. et al. (2007) Phenotypic characterization of human colorectal cancer stem cells. Proc. Natl. Acad. Sci. U.S.A. 104: 10158-10163

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