Pe conjugated epcam antibody

Manufactured by Cell Signaling Technology

The PE-conjugated EpCAM antibody is a laboratory reagent used to detect and analyze the expression of the Epithelial Cell Adhesion Molecule (EpCAM) on the surface of cells. The antibody is conjugated with the fluorescent dye Phycoerythrin (PE), which allows for the identification and quantification of EpCAM-positive cells using flow cytometry or other fluorescence-based techniques.

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Lab products found in correlation

Draq5, by Cell Signaling Technology (2 mentions) Pe cf594 conjugated cd45 antibody, by BD (2 mentions) Cellevent caspase 3 7 green detection reagent, by Thermo Fisher Scientific (2 mentions) Pe conjugated cd66b antibody, by STEMCELL (1 mentions) Alexa fluor 488 conjugated cd11b antibody, by STEMCELL (1 mentions) Clone g10f5, by STEMCELL (1 mentions) Calcein blue am, by Thermo Fisher Scientific (1 mentions) Clone hi30, by BD (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions)

2 protocols using pe conjugated epcam antibody

Multiparameter Imaging Flow Cytometry

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Cell viability, leukocyte activation, and platelet–leukocyte adhesion were studied using the ImageStream^X Mark II imaging flow cytometer (Amnis Corporation) equipped with a 40× objective, 6 imaging channels, and 405, 488, and 642 nm lasers. Whole blood was diluted 1:33 in RPMI 1640 medium (supplemented with 10 mM HEPES; Life Technologies) and stained with the following where applicable: calcein blue, AM (10 µM; Life Technologies); Pacific Blue-conjugated CD41 antibody (1:100; clone HIP8; BioLegend, Cat# 303713); CellEvent Caspase-3/7 Green Detection Reagent (5 µM; Life Technologies); Alexa Fluor 488-conjugated CD11b antibody (1:500; clone ICRF44; Stemcell Technologies, Cat# 60040AD); PE-conjugated CD66b antibody (1:150; clone G10F5; Stemcell Technologies, Cat# 60086PE); PE-conjugated EpCAM antibody (1:250; clone VU1D9; Cell Signaling Technology, Cat# 8995s); PE-CF594-conjugated CD45 antibody (1:666; clone HI30; BD Biosciences, Cat# 562279); and DRAQ5 (1 μM; Cell Signaling Technology). Single cells were gated using the nuclear stain DRAQ5. Viable cells were defined as calcein-positive but caspase-3/7-negative.

Wong K.H., Tessier S.N., Miyamoto D.T., Miller K.L., Bookstaver L.D., Carey T.R., Stannard C.J., Thapar V., Tai E.C., Vo K.D., Emmons E.S., Pleskow H.M., Sandlin R.D., Sequist L.V., Ting D.T., Haber D.A., Maheswaran S., Stott S.L, & Toner M. (2017). Whole blood stabilization for the microfluidic isolation and molecular characterization of circulating tumor cells. Nature Communications, 8, 1733.

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Imaging Flow Cytometry for CTC Analysis

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Imaging flow cytometry was performed using the ImageStream^X Mark II imaging flow cytometer (Amnis Corporation) equipped with a 40× objective, six imaging channels, and 405, 488, and 642 nm lasers. For analysis of cell viability and EpCAM expression, Brx cells obtained from culture or captured/released from our microfluidic device were resuspended in RPMI (supplemented with 0.3% BSA and HEPES) and stained with the following antibodies and stains where applicable: calcein blue AM (2.5 μM, ThermoFisher Scientific), CellEvent caspase 3/7 green detection reagent (1.75 μM, Life Technologies), PE-conjugated EpCAM antibody (1:260, clone VU1D9, Cell Signaling), PE-cf594-conjugated CD45 antibody (1:400, clone H130, BD Bioscience), and DRAQ5 (1 μM, Cell Signaling Technologies). Cells were acquired/gated using the nuclear marker DRAQ5. EpCAM positive cells were gated for viable cells (calcein positive and caspase 3/7 negative) versus dead cells (caspase 3/7 positive).

Park M.H., Reátegui E., Li W., Tessier S.N., Wong K.H., Jensen A.E., Thapar V., Ting D., Toner M., Stott S.L, & Hammond P.T. (2017). Enhanced Isolation and Release of Circulating Tumor Cells Using Nanoparticle Binding and Ligand Exchange in a Microfluidic Chip. Journal of the American Chemical Society, 139(7), 2741-2749.

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