Clone:
REAL546
Type of antibody:
Releasable antibodies, Primary antibodies, Recombinant antibodies
Applications:
FC
Alternative names:
EGP40, Ep-CAM, KSA, TROP1, MK-1

Extended validation for CD326 (EpCAM) 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 REAL546
REA764++
HEA-125++
CO17-1A++
Cells were incubated with an excess of purified unconjugated CD326 (EpCAM) (REAL546) 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 CD326 (EpCAM). SK-BR-3 cells were stained with CD326 (EpCAM) antibodies and plotted against the side scatter. As a control, CD326 (EpCAM) 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 CD326 (EpCAM). SK-BR-3 cells were stained with CD326 (EpCAM) antibodies and plotted against the side scatter. As a control, CD326 (EpCAM) 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 CD326 (EpCAM). SK-BR-3 cells were stained with CD326 (EpCAM) antibodies and plotted against the side scatter. As a control, CD326 (EpCAM) 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 CD326 (EpCAM). SK-BR-3 cells were stained with CD326 (EpCAM) antibodies and plotted against the side scatter. As a control, CD326 (EpCAM) 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 CD326 (EpCAM). SK-BR-3 cells were stained with CD326 (EpCAM) antibodies and plotted against the side scatter. As a control, CD326 (EpCAM) 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 CD326 (EpCAM). SK-BR-3 cells were stained with CD326 (EpCAM) antibodies and plotted against the side scatter. As a control, CD326 (EpCAM) 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 CD326 (EpCAM). SK-BR-3 cells were stained with CD326 (EpCAM) antibodies and plotted against the side scatter. As a control, CD326 (EpCAM) 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 CD326 (EpCAM) (REAL546). 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 CD326 (EpCAM) (REAL546). 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 CD326 (EpCAM) (REAL546). 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 CD326 (EpCAM) Antibody, anti-human,
REAlease
®

Overview

Clone REAL546 is an antibody fragment derived from the full CD326 (EpCAM) 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 REAL546 recognizes CD326, also known as human epithelial antigen (HEA), epithelial cell adhesion molecule (EpCAM), or epithelial-specific antigen (ESA), and is involved in cell adhesion. The CD326 antigen is broadly expressed on the basolateral surface of carcinoma and epithelial cells in tissues or on circulating tumor cells and cancer stem cells, but is not found on melanoma, neuroblastoma, sarcoma, lymphoma, leukemia cells, or normal fibroblasts. Furthermore, it is suggested, that CD326 could be used as a surface marker for human ESCs.
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

EGP40, Ep-CAM, KSA, TROP1, MK-1

Detailed product information

Technical specifications

CloneREAL546
Clonalitymonoclonal
Isotype controlControl Antibody
Hostcell line
Type of antibodyReleasable antibodies, Primary antibodies, Recombinant antibodies
Specieshuman
AntigenCD326 (EpCAM)
Alternative names of antigenEGP40, Ep-CAM, KSA, TROP1, MK-1
Distribution of antigenepithelial cells, cancer stem cells, ES and iPS cells, lung
RRIDAB_2801849, AB_2801852, AB_2801851, AB_2801855, AB_2801854, AB_2801858, AB_2801857, AB_2811704, AB_2811705, AB_2811697, AB_2811706

Resources for CD326 (EpCAM) Antibody, anti-human,
REAlease
®

Certificates

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

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REAlease
®

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