Protein samples of cell lysate or Co-IP product were subjected to Western blot analysis as previously described [38]. The primary antibodies used in this study were rabbit anti-OSBP (Proteintech Group, Inc., Rosemont, IL), mouse anti-GAPDH (Santa Cruz Biotechnology, Inc., Dallas, TX), rabb...
primary antibodies at 4°C overnight, the membranes were washed 5 times in 0.1% TBS-Tween and incubated for 1 h with a chicken anti-rabbit IgG horseradish peroxidase-conjugated secondary antibody (dilution, 1:2000; cat. no. sc-516,087; Santa Cruz Biotechnology, Inc., Dallas, TX, USA)...
Antibodies used were MDM2 (1:1:1 mix of mouse monoclonal 4B2, 2A9, 4B11 hybridoma supernatant), p53 (1:1:1 mix of mouse monoclonal 240,421,1801 hybridoma supernatant), and MDMX (Proteintech, Rosemont, IL, USA), actin-HRP (Santa Cruz Biotechnology, Dallas, TX, USA), E-cadherin (Cell...
Recall Th1 responses were examined by stimulating PBMC with 1:100-diluted recall stimuli Candida albicans (Allermed Laboratories, San Diego, CA, USA) and tetanus toxoid (Santa Cruz Biotechnology, Dallas, TX, USA). An empiric method of determining anti-HER2 Th1 response specificity was employed [...
After 24 h, cells were incubated in SFM containing 100 μM of the exonuclease inhibitor ARL-67156 (Santa Cruz Biotechnologies, Dallas, TX, USA) for 30 min at 37 °C. Then, the cells were stimulated with 2 µg of Thy-1-Fc per well, for 10 min. Where indicated, cells were ...
The antibody against vimentin was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The antibody against glyceraldehyde 3-phosphate dehydrogenase (GAPDH; loading control) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Transwell assays Cell invasion and migration were measured using...
Children’s Research Institute Mouse Genome Engineering Core, University of Texas Southwestern Medical Center, Dallas, TX, 75390, USA Lin Li, Yu Zhang & Hao Zhu Manchester Collaborative Centre for Inflammation Research (MCCIR), School of Biological Sciences, Faculty of Biology, Medicine and Health,...
For IHC, spheroids were cultured for seven days and then harvested and fixed in 4% paraformaldehyde (Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4 °C. Thereafter, the cells were rinsed with PBS, stained with 0.1% toluidine blue D (Merck, Kenilworth, NJ, USA) and centrifuged...
(1:100, v/v, Santa Cruz Biotechnology, Dallas, TX, USA) or anti-CD8 (1:100, Santa Cruz Biotechnology, Dallas, TX, USA) antibodies in a humidified chamber overnight at 37 °C. Sections were washed with 1× PBS and incubated with secondary antibody FITC-conjugated anti-mouse Alexa Fluor...
FOXA2 for endoderm (#07-633; Millipore), beta III tubulin (#ab41489; Abcam) for ectoderm, and CD31 (#sc-1506; Santa Cruz Biotechnology, Dallas, TX, USA) for mesoderm were used to stain the EB-derived cells. Teratoma formation and in vivo differentiation of induced pluripotent stem cells...