WebApr 12, 2024 · The cell proliferative ability was assessed via Cell Counting Kit‑8 assay and using 5‑ethynyl‑2'‑deoxyuridine staining. Flow cytometry and western blot analyses were applied for the assessment of cell apoptosis. Cell autophagy was investigated by employing western blot analysis and immunofluorescence staining. WebNov 12, 2024 · The In-Cell Western (ICW) assay is based on the quantification of immunofluorescence signals from proteins in fixed cultured cells using target-specific primary antibodies. Gene expression levels, phosphorylation of proteins, caspases activity, cell proliferation assay and viral titration represent the common application of this assay [ …
High-Throughput Characterization of Viral and Cellular Protein ... - PubMed
WebThe cells are trypsinized and then seeded into a 96-well tissue culture plate. Black-walled plates may be used to reduce side-scatter. 6The recommended seeding density is 0.2 x 10 cells per mL. Allow the cells to grow to confluence (for Capan-2 cells approximately 48 hours). Since alpha tubulin is cytosolic, the cells are fixed and permeabilized. WebWestern Blot Antibody Dilution Calculator. Use this calculator to help you correctly dilute your antibodies for western blotting. To correctly dilute secondary antibodies for western blotting it is recommended to first make an antibody working stock solution of 1 µg/mL and dilute further for the final antibody incubation solution. easy beer batter hush puppies recipes
Quantitative Analysis of Signal Transduction with In-Cell Western ...
WebApr 13, 2024 · The cultured cells were lysed, and the whole-cell extracts were assayed via Western blotting using antibodies against NLRP3 and the active form of caspase-1, following the protocol previously described. Additionally, the cultures were pre-treated with caspase-1 inhibitor (Z-YVAD-fmk 1 and 10 μg/mL) for 30 min, and stimulated with IL-17A … WebThe In-Cell Western TM (ICW) Assay is a quantitative immunofluorescence assay that combines Western blotting with high-throughput ELISA. Using this technique, researchers … WebMTAP-wt cells after 72 hr of treatment with DMSO control or various doses of AG-270, measured by in-cell western DMSO = dimethyl sulfoxide Figure 6. T reatment with AG-270 induced DNA damage and mitotic defects in MTAP-null cells • AG-270 demonstrated an additive to synergistic effect, as defi ned by Chou and Talalay,8 when cuny graduate center bursar office