wash buffer  (Santa Cruz Biotechnology)

Journal: Cell Death & Disease

Article Title: Cell surface galectin-3 defines a subset of chemoresistant gastrointestinal tumor-initiating cancer cells with heightened stem cell characteristics

doi: 10.1038/cddis.2016.239

Figure Lengend Snippet: Surface Gal3 defines a subtype of epithelial CSC. ( a ) Colorectal cancer cells LiM6 and DLD1 were investigated by flow cytometry for cell surface markers CD24/CD44 (left). CD24+/CD44+-cells (in black) were then investigated for CD166/EpCAM-expression (middle) and CD24+/CD44+/CD166+/EpCAM+-positive cells (in black middle) for Gal3-expression (right panel), where ++++ Gal3 − in green refers to cells positive for CD24, CD44, CD166, and EpCAM, but negative for Gal3, and ++++ Gal3 + in red refers to cells positive for CD24, CD44, CD166, EpCAM, and Gal3. ( b ) Pancreatic cancer cell lines AsPC1 and L3.6pl were subjected to the same analysis as in ( a )

Article Snippet: To identify and isolate CSC, parental cells were collected in Versene solution (Invitrogen, Carlsbad, CA, USA, cat#15040), washed and blocked with flow cytometry buffer (Santa Cruz Biotechnology, Dallas, TX, USA), labeled with the following primary conjugated antibodies and their matched isotypes: anti-CD24-PE-Cy7(561646), anti-EpCAM-FITC (BD Bioscience, San Jose, CA, USA, 347197) anti-CD44-APC-eFluor 780(47044182), anti-CD166-PerCPeFluor 710(46166842), anti Gal3-PE(12-5301-83) (all from - eBioscience, San Diego, CA, USA).

Techniques: Flow Cytometry, Expressing

Journal: Cell Death & Disease

Article Title: Cell surface galectin-3 defines a subset of chemoresistant gastrointestinal tumor-initiating cancer cells with heightened stem cell characteristics

doi: 10.1038/cddis.2016.239

Figure Lengend Snippet: Silencing of Gal3 shifts CSC subset to a Gal3 negative CSC subset. ( a ) Western blot analysis for comparison of total Gal3 in LiM6TR or DLD1TR colon cancer cells and L3.6pl or AsPC1 pancreatic cancer cells after infection with lentiviral particles for control-shRNA ( c ) or Gal3-shRNA (G). Each pair of control and Gal3 knockdown cells were run side by side on the same gel. Comparison of Gal3 expression between cell lines has been previously published ( b ) Flow cytometric analysis of colon cancer and pancreatic cancer cells after infection with lentiviral particles for control-shRNA (red trace) or Gal3-shRNA (green trace). Black trace is background staining. ( c ) ALDH activity was analyzed by flow cytometry. ALDEFLUOR activity in colon cancer and pancreatic cancer cells after infection with lentiviral particles for control-shRNA (blue trace) or Gal3-shRNA (green trace). Gray profile is background staining

Article Snippet: To identify and isolate CSC, parental cells were collected in Versene solution (Invitrogen, Carlsbad, CA, USA, cat#15040), washed and blocked with flow cytometry buffer (Santa Cruz Biotechnology, Dallas, TX, USA), labeled with the following primary conjugated antibodies and their matched isotypes: anti-CD24-PE-Cy7(561646), anti-EpCAM-FITC (BD Bioscience, San Jose, CA, USA, 347197) anti-CD44-APC-eFluor 780(47044182), anti-CD166-PerCPeFluor 710(46166842), anti Gal3-PE(12-5301-83) (all from - eBioscience, San Diego, CA, USA).

Techniques: Western Blot, Comparison, Infection, Control, shRNA, Knockdown, Expressing, Staining, Activity Assay, Flow Cytometry