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Apc conjugated anti epcam

Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany

The APC-conjugated anti-EpCAM is a fluorochrome-labeled antibody that binds to the Epithelial Cell Adhesion Molecule (EpCAM) protein. EpCAM is expressed on the surface of epithelial cells and cancer cells of epithelial origin. The APC (Allophycocyanin) fluorochrome enables the detection and identification of EpCAM-positive cells using flow cytometry or other fluorescence-based techniques.

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3 protocols using apc conjugated anti epcam

1

Characterizing Liver Cell Populations by Flow Cytometry

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For the determination of LPC percentages within liver NPCs, NPCs were incubated for 30 min on ice with the following antibodies: APC-conjugated anti-EpCAM and FITC-conjugated anti-CD45 (eBioscience). For the determination of the subtypes of liver leukocytes, liver leukocytes were incubated for 30 min on ice with the following antibodies: APC/FITC-conjugated anti-Ly6G, PE-conjugated anti-CD11b and FITC/APC-conjugated anti-CD45; FITC/PE-conjugated anti-CD8, PE-conjugated anti-CD4 and APC-conjugated anti-CD45 (eBioscience, San Diego, CA, USA). For the characterization of LPCs cultured in vitro, LPCs were trypsinized and incubated for 30 min on ice with the following antibodies: APC-conjugated anti-EpCAM, PE-conjugated anti-CD44, APC-conjugated anti-CD49f or FITC-conjugated anti-CD45 (eBioscience). For the detection of ROS, liver leukocytes were incubated with 2-[6-(4′-amino)phenoxy-3H-xanthen-3-on9-yl]benzoic acid (APF; Invitrogen, Carlsbad, CA, USA) fluorescence as reported before [26 (link)]. FCM measurements were performed with Aria II (BD Biosciences). Data were analyzed using FlowJo software (Tree Star, San Carlos, CA, USA).
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2

Isolation and Culture of Liver Progenitor Cells

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LPCs were isolated as described previously [24 (link)]. Briefly, liver tissues were removed from CDE diet-fed mice after in-situ perfusion using a two-step collagenase perfusion method and then were incubated with 1 mg/ml collagenase D (Roche) and 1 mg/ml pronase (Roche) at 37 °C for 30 min. Non-parenchymal cells (NPCs) were separated from hepatocytes by repeated low-speed centrifugation. Next, NPCs were collected and suspended in a discontinuous gradient of 20% and 50% Percoll (GE Healthcare, Pittsburgh, PA, USA) and centrifuged continuously at 1400 × g for 20 min to enrich LPCs. The enriched LPCs were labelled with APC-conjugated anti-EpCAM and FITC-conjugated anti-CD45 (eBioscience, San Diego, CA, USA) antibodies, and EpCAM+CD45 cells were isolated by fluorescence-activated cell sorting. These LPCs were cultured in type I collagen-coated dishes (BD Biosciences, San Jose, CA, USA). The standard culture medium for LPCs was DMEM/F12 (Gibco, Grand Island, NY, USA) supplemented with 10% foetal bovine serum, 1 μg/ml insulin (Sigma-Aldrich), 1 × 10− 7 mol/L dexamethasone (Sigma-Aldrich), 1% penicillin/streptomycin (Invitrogen), 50 ng/ml hepatocyte growth factor (PeproTech, Rocky Hill, NJ, USA), 20 ng/ml epidermal growth factor (PeproTech), 20 ng/ml FGF (PeproTech) and 1× insulin–Transferrin–Selenium–Ethanolamine (Invitrogen).
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3

Psoriasis Induction and Analysis Protocol

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Aldara ® (IMQ) 5% cream (MEDA Pharmaceuticals, Vienna, Austria) was used for inducing psoriatic skin changes in the mice. 1% rapamycin (Merck Calbiochem, Darmstadt, Germany) ointment was made by mixing 50 mg petroleum jelly with 10 μl of the corresponding rapamycin stock, 50 mg/ml. P-mTOR S2448 #2976 and P-S6 S235/6 #2211 antibodies were from Cell Signaling Technology, Frankfurt, Germany. Keratin 6 antibody (Ks6. KA12) from Thermo Scientific, Darmstadt, Germany and caspase-14 antibody (NB100-56126) was from Novus Biologicals, Wiesbaden, Germany. NIMP-R14 Ly-6G/C (ab2557) was from Abcam, Cambridge, UK. Fluorescein isothiocyanate (FITC)conjugated anti-CD3, phycoerythrin (PE)-conjugated anti-CD4, PE-conjugated anti-CD11c, allophycocyanin (APC)-conjugated anti-CD11b, FITC-conjugated anti-F4/80, APC-conjugated anti-B220, PE-conjugated anti-langerin, APC-conjugated anti-EpCAM all were from eBioscience, Darmstadt, Germany, while peridinin chlorophyll protein complex (PerCP/Cy5)-conjugated anti-Siglec-H was from Biolegend, Koblenz, Germany and PerCP-conjugated anti-CD8 was purchased from BD Bioscience, Heidelberg Germany.
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