TOV112D and Saos-2 cells harboring various p53 mutants were treated with vehicle (DMSO) or 30μM UCI-LC0023 for 3 hours. For fractionation assay with Crm1 inhibitor KPT-185 (Selleckchem, S7125), cells were treated with 25 nM KPT-185 for 24 hours. Cell pellets were harvested and washed with ice-cold PBS. The nuclear and chromatin fractions were prepared as previously described (Mendez and Stillman, 2000 ). Equivalent fractions of proteins from the soluble nuclear extract and chromatin bound nuclear extract were resolved on SDS-PAGE and transferred on to PVDF membranes. The blots were probed with p53 (FL-393) and Histone H3 (marker for chromatin fraction) antibodies (Abcam, ab1791).
Kpt 185
Manufactured by Selleck Chemicals
Sourced in Germany
KPT-185 is a laboratory instrument used for the analysis and purification of chemical compounds. It is designed to perform chromatographic separations, a common technique in analytical chemistry and organic synthesis. The core function of KPT-185 is to facilitate the isolation and purification of target compounds from complex mixtures.
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Lab products found in correlation
3 protocols using kpt 185
1
Analyzing Mutant p53 in TOV112D and Saos-2 Cells
2
Assessing Cell Viability and Proliferation
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Cells were seeded at 0.8 × 106 cells/ml for 24 h. Next, cells were treated with 1 μM KPT-185 (Selleck Chemicals, Munich, Germany) or 20 nM LMB (Enzo Life Sciences, Brussels, Belgium). Cell viability and proliferation were assessed by (i) Trypan Blue exclusion dye assay, (ii) the cell proliferation reagent WST-1 (Roche Life Sciences) according to the manufacturer’s instructions, and (iii) flow cytometry using FITC-Ki-67 (BD Biosciences, CA, USA). The detailed protocols are described in the Online Supplementary Methods.
3
Assessing Cell Viability and Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded at 0.8x10 6 cells/ml for 24 h. Next, cells were treated with 1 µM KPT-185 (Selleck Chemicals, Munich, Germany) or 20 nM LMB (Enzo Life Sciences, Brussels, Belgium). Cell viability and proliferation was assessed by (i) Trypan Blue exclusion dye assay, (ii) the cell proliferation reagent WST-1 (Roche Life Sciences) according to the manufacturer's instructions, and (iii) flow cytometry using FITC-Ki-67 (BD Biosciences, CA, USA). The detailed protocols are described in the Online Supplementary Methods.
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