Clone:
REA664
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
CD44s, EMCR III, H-CA, Pgp-1, HERMES, Ly-24, HUTCH-I, Hermes antigen

Extended validation for CD44 Antibody, anti-mouse, 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 REA664
IM7++
IM7.8.1++
Cells were incubated with an excess of purified unconjugated CD44 (REA664) 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. Splenocytes from C57BL/6 mice 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. Splenocytes from C57BL/6 mice 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. Splenocytes from C57BL/6 mice 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. Splenocytes from C57BL/6 mice 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 (REA664). 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 (REA664). 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 (REA664). 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-mouse, REAfinity™

Overview

Clone REA664 recognizes the mouse CD44 antigen, which is also known as Pgp-1, HCELL, HUTCH-I, HCAM, HERMES, or Ly-24. CD44 is a 80–95 kDa membrane glycoprotein and expressed by a variety of hematopoetic and non-hematopoetic cells. Different isoforms are generated by alternative splicing. Clone REA664 reacts with all isoforms and both allelic forms of CD44. CD44 functions as a receptor for hyaluronic acid (HA) and mediates cell-cell and cell-matrix interactions through its affinity for HA, and possibly also through its affinity for other ligands, such as osteopontin, collagens, and matrix metalloproteinases (MMPs). The interaction between galectin-9 and CD44 increases the stability and function of adaptive regulatory T cells. CD44 expression is upregulated on naive T cells upon activation and high CD44 expression is maintained on memory T cells. Therefore, CD44 is often used to discriminate CD44
high
antigen–experienced T cells from CD44
low
(CD62L
+
) naive T cells.
Additional information: Clone REA664 displays negligible binding to Fc receptors.

Alternative names

CD44s, EMCR III, H-CA, Pgp-1, HERMES, Ly-24, HUTCH-I, Hermes antigen

Detailed product information

Technical specifications

CloneREA664
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
,
cynomolgus monkey (
Macaca fascicularis
)
,
chimpanzee (
Pan troglodytes
)
, baboon, human, pig, cat, dog, cow, horse
AntigenCD44
Alternative names of antigenCD44s, EMCR III, H-CA, Pgp-1, HERMES, Ly-24, HUTCH-I, Hermes antigen
Molecular mass of antigen [kDa]83
Distribution of antigenendothelial cells, epithelial cells, T cells, red blood cells, lymphocytes, CNS cells, mesenchymal stem cells, plasma cells, ES and iPS cells, cancer stem cells, bone marrow, kidney, skeletal muscle, skin
Entrez Gene ID12505
RRIDAB_2658157, AB_2658160, AB_2658161, AB_2751543, AB_2751628, AB_2733233, AB_2733234, AB_2733120, AB_2752051, AB_2752040, AB_2751557

References for CD44 Antibody, anti-mouse, REAfinity™

Publications

  1. Ponta, H. et al. (2003) CD44: From adhesion molecules to signalling regulators. Nat. Rev. Mol. Cell Biol. 4(1): 33-45
  2. Wu, C. et al. (2014) Galectin-9-CD44 interaction enhances stability and function of adaptive regulatory T cells. Immunity 41(2): 270-282
  3. He, Q. et al. (1992) Molecular isoforms of murine CD44 and evidence that the membrane proximal domain is not critical for hyaluronate recognition. J. Cell Biol. 119(6): 1711-1719

Related products for
CD44 Antibody, anti-mouse, REAfinity™

2 products available