Clone:
REA199
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
CDH5, 7B4

Extended validation for CD144 (VE-Cadherin) 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 REA199
55-7H1++
16B1-
BV9n/a
Cells were incubated with an excess of purified unconjugated CD144 (VE-Cadherin) (REA199) 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 CD144 (VE-Cadherin). Huvec cells were stained with CD144 (VE-Cadherin) antibodies and with a suitable counterstaining. As a control, CD144 (VE-Cadherin) 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. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD144 (VE-Cadherin). Huvec cells were stained with CD144 (VE-Cadherin) antibodies and with a suitable counterstaining. As a control, CD144 (VE-Cadherin) 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. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD144 (VE-Cadherin). Huvec cells were stained with CD144 (VE-Cadherin) antibodies and with a suitable counterstaining. As a control, CD144 (VE-Cadherin) 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. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD144 (VE-Cadherin). Huvec cells were stained with CD144 (VE-Cadherin) antibodies and with a suitable counterstaining. As a control, CD144 (VE-Cadherin) 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. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD144 (VE-Cadherin). Huvec cells were stained with CD144 (VE-Cadherin) antibodies and with a suitable counterstaining. As a control, CD144 (VE-Cadherin) 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. 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 CD144 (VE-Cadherin) (REA199). 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 CD144 (VE-Cadherin) (REA199). 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 CD144 (VE-Cadherin) (REA199). 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 CD144 (VE-Cadherin) Antibody, anti-human, REAfinity™

Overview

Clone REA199 recognizes CD144 (VE-Cadherin), a 120 KDa type II cadherin. Cadherins are cell adhesion molecules and mediate Ca
2+
dependent homophilic interactions. CD144 contains five extracellular cadherin (EC) domains and like other cadherins can interact directly via its C-terminus with cytoplasmic proteins such as β-catenin, plaktoglobin, and p120. Plaktoglobin und β-catenin bind to α-catenin, which in turn interacts with several actin-binding proteins, α-actinin, ajuba, zonula occludens-1 (ZO-1). Further indirect interactions of CD144 with partners such as SHP-2, VEGFR-2, Csk, and PAR-3, 6 allows CD144 to not only regulate the stability and strength of cell adhesion but also to serve functions such as sensing of shear forces, anti-proliferative, and anti-apoptotic effects.
Additional information: Clone REA199 displays negligible binding to Fc receptors.

Alternative names

CDH5, 7B4

Detailed product information

Technical specifications

CloneREA199
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD144 (VE-Cadherin)
Alternative names of antigenCDH5, 7B4
Molecular mass of antigen [kDa]120
RRIDAB_2751492, AB_2819545, AB_2819510, AB_2857828, AB_2857821, AB_2655156, AB_2655157, AB_2655158, AB_2655159, AB_2655162, AB_2655163, AB_2751528

Resources for CD144 (VE-Cadherin) Antibody, anti-human, REAfinity™

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for CD144 (VE-Cadherin) Antibody, anti-human, REAfinity™

Publications

  1. Vincent, P. A. et al. (2004) VE-cadherin: adhesion at arm's length. Am. J. Physiol., Cell Physiol. 286: C987-C997
  2. Nelson, C. M. and Chen, C. S. (2003) VE-cadherin simultaneously stimulates and inhibits cell proliferation by altering cytoskeletal structure and tension. J. Cell. Sci. 116: 3571-3581
  3. Odell, A. F. et al. (2012) A VE-cadherin–PAR3–α-catenin complex regulates the Golgi localization and activity of cytosolic phospholipase A2α in endothelial cells. Mol. Biol. Cell 23: 1783-1796

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