Journal: STAR Protocols

Article Title: Protocol for quantifying drug sensitivity in 3D patient-derived ovarian cancer models

doi: 10.1016/j.xpro.2024.103274

Figure Lengend Snippet:

Article Snippet: CD326 (EpCAM) antibody, anti-human, PE, REAfinity (flow cytometry 1:200) , Miltenyi Biotec , Cat# 130-110-999; RRID: AB_2657495.

Techniques: Immunofluorescence, Flow Cytometry, Control, Recombinant, Red Blood Cell Lysis, Saline, Software, Cell Culture, Plasmid Preparation, Microscopy

Journal: Journal of Breast Cancer

Article Title: The Presence of EpCAM - /CD49f + Cells in Breast Cancer Is Associated with a Poor Clinical Outcome

doi: 10.4048/jbc.2015.18.3.242

Figure Lengend Snippet: Dual-immunohistochemistry (IHC) and immunofluorescence (IF) staining on tissue microarray (TMA) slides. Dual-IHC and IF staining with primary antibodies of EpCAM and CD49f were performed on TMA tissue. (A) Patients only contain EpCAM-positive cancer cells (membrane); (B) Patient show only CD49f-positive cancer cells (cytoplasm); (C) Case express none or both (D) biomarkers. In IF staining (E-P), green signal shows the expression of CD49f, red signal shows EpCAM with DAPI staining for nuclei. (E-H) One case that tumor cell expression only EpCAM on the membrane (white arrows); (I-L) One case that tumor cell express only CD49f (red arrow); (M-P) One case with tumor cells express both (yellow arrow) (for both IHC and IF, magnification is actually ×400).

Article Snippet: IHC was performed using antibodies against EpCAM (TA310957, 1:300; OriGene, Rockville, USA) and CD49f (TA506627, 1:150; OriGene) with an Autostainer with the EnVision G|2 Double Stain System ((DAKO company, Code K5361, Glostrup, Denmark) Rabbit/Mouse (DAB+/Permanent Red) according to the manufacturer's instructions.

Techniques: Immunohistochemistry, Immunofluorescence, Staining, Microarray, Membrane, Expressing

Journal: Journal of Breast Cancer

Article Title: The Presence of EpCAM - /CD49f + Cells in Breast Cancer Is Associated with a Poor Clinical Outcome

doi: 10.4048/jbc.2015.18.3.242

Figure Lengend Snippet: Prevalence of EpCAM - /CD49f + with histopathologic characteristics. Analyzing of 150 breast cancer samples stratified to tumor size (A), node status (B), tumor grade (C; low=grade I and II, high=grade III), recurrence (D), distant metastasis (E), and estrogen receptor (ER) expression (F). Tumor containing EpCAM - /CD49f + are associated with higher tumor grade (C; p =0.031) and distant metastasis (E; p =0.048). Chi-square exact and Fisher exact test were performed.

Article Snippet: IHC was performed using antibodies against EpCAM (TA310957, 1:300; OriGene, Rockville, USA) and CD49f (TA506627, 1:150; OriGene) with an Autostainer with the EnVision G|2 Double Stain System ((DAKO company, Code K5361, Glostrup, Denmark) Rabbit/Mouse (DAB+/Permanent Red) according to the manufacturer's instructions.

Techniques: Expressing

Journal: Journal of Breast Cancer

Article Title: The Presence of EpCAM - /CD49f + Cells in Breast Cancer Is Associated with a Poor Clinical Outcome

doi: 10.4048/jbc.2015.18.3.242

Figure Lengend Snippet: Univariate survival analysis of EpCAM - /CD49f + with disease-free survival (DFS) and overall survival (OS). All the patients' tumor samples were stained with EpCAM and CD49f. (A, B) The present of EpCAM - /CD49f + tumor cells have significant negative association with both DFS and OS (EpCAM - /CD49f + : p DFS =0.009; p OS =0.001).

Article Snippet: IHC was performed using antibodies against EpCAM (TA310957, 1:300; OriGene, Rockville, USA) and CD49f (TA506627, 1:150; OriGene) with an Autostainer with the EnVision G|2 Double Stain System ((DAKO company, Code K5361, Glostrup, Denmark) Rabbit/Mouse (DAB+/Permanent Red) according to the manufacturer's instructions.

Techniques: Staining

Journal: Nanoscale

Article Title: Multi-layer assembly of cellulose nanofibrils in a microfluidic device for the selective capture and release of viable tumor cells from whole blood.

doi: 10.1039/d0nr05375a

Figure Lengend Snippet: Figure 1 Schematics of LbL CNF assembly for affinity cell capture and enzymatic release and AFM characterization. A- CNF LbL buildup at neutral pH on a surface modified with PAH. Both PEI and PAH have a global positive charge, while CNF has a global negative charge. B- Affinity cell capture after conjugating anti-EpCAM antibodies on the upper layer of CNF (top), followed by digestion of the CNF layers using a cocktail of cellulolytic enzymes resulting in the release of trapped cells (bottom). C to E – AFM characterization of 1, 3 or 5 CNF layers on a silicon wafer (10x10 μm). Scale bars refer to a width of 1 μm. F – Height profile of the CNF LbL assembly for each of the tested number of layers. Layer 0 corresponds to the profile of the silicon wafer after scratching the CNF LbL assembly from the surface.

Article Snippet: Anti-EpCAM mouse monoclonal antibodies (mAbs) purchased from R&D Systems (MAB960, clone #158210) were grafted on the cellulose via carboxymethyl groups via EDC-NHS chemistry.

Techniques: Modification

Journal: Nanoscale

Article Title: Multi-layer assembly of cellulose nanofibrils in a microfluidic device for the selective capture and release of viable tumor cells from whole blood.

doi: 10.1039/d0nr05375a

Figure Lengend Snippet: Figure 3 Effect of increasing numbers of CNF layers on CNF surface density, antibody immobilization, enzymatic CNF release and cell capture/release. Dotted lines indicate the border of the microchannel. The plots superimposed with the fluorescence microscopy images quantify the average lateral fluorescent intensity across the channel. A- Cellulose surface density and homogeneity measured with Carbotrace™ 680 after coating 1,3 or 5 CNF layers, from left to right, respectively. B- Alexa 430 labeled anti-EpCAM antibody surface density with 1,3 or 5 CNF layers from left to right, respectively. The results in A and B were measured in 3 independent devices, each coated with 1, 3 or 5 CNF layers. C- Cellulose after coating (left) and after 30 min of enzyme digestion for 1 (top) and 5 (bottom) layers of CNF. The values indicate the background subtracted average fluorescence on the channel surface. D- HCT 116 cell capture (left) and subsequent release (right) for 1 (top) and 5 (bottom) layers of CNF. The values indicate the total cell count in each image.

Article Snippet: Anti-EpCAM mouse monoclonal antibodies (mAbs) purchased from R&D Systems (MAB960, clone #158210) were grafted on the cellulose via carboxymethyl groups via EDC-NHS chemistry.

Techniques: Fluorescence, Microscopy, Labeling, Cell Counting

Journal: PLoS Pathogens

Article Title: Receptor-Targeted Nipah Virus Glycoproteins Improve Cell-Type Selective Gene Delivery and Reveal a Preference for Membrane-Proximal Cell Attachment

doi: 10.1371/journal.ppat.1005641

Figure Lengend Snippet: ( A ) Cell entry is EpCAM dependent but independent of ephrin-B2. Representative flow cytometry plots out of three independent experiments of CHO-K1, CHO-EpCAM, CHO-ephrin-B2 cells, and of a mixed culture composed of CHO-EpCAM and CHO-ephrin-B2 (1:1 ratio) monitored 72 h after transduction with NiVmut EpCAM -LV, NiVwt-LV or VSV-LV (MOI of 1). EpCAM expression was detected by an APC-coupled human EpCAM specific antibody. ( B ) To ascertain stability of transduction with the EpCAM-targeted vector, CHO-EpCAM cells were cultivated for further 30 days after transduction with the indicated MOIs. The percentage of GFP-positive cells was determined by flow cytometry at the indicated time points. One representative out of three independent experiments is shown.

Article Snippet: Human EpCAM was detected by an Allophycocyanin (APC) labeled mouse anti-EpCAM antibody (clone HEA-125, Miltenyi Biotec, Bergisch Gladbach, Germany, dilution 1:100).

Techniques: Flow Cytometry, Transduction, Expressing, Plasmid Preparation