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This protocol outlines the extraction of proteins from neuronal cancer cells. The cells are washed with a buffer and lysed via enzymatic and mechanical disruption. The extracted protein-rich supernatant is collected after centrifugation.
1. Cell Line Preparation
2. Protein Expression Validation
Name | Company | Catalog Number | Comments |
Mouse Motor Neuron-Like hybrid Cell Line (NSC-34) | tebu-bio | CLU140-A | |
DMEM 4.5g/L L-glucose | Biological Industries | 01-055-1A | |
FBS European Grade | Biological Industries | 04-007-1A | |
SL 16R Centrifuge | Thermo Scientific | 75004030 | |
Thermo Forma Series ii Water Jacketed CO2 Incubator | Thermo Scientific | 3111 | |
Cover glass 12mm dia. thick 0.13-0.17mm | Bar Naor LTD | ||
Axiovert 100 inverted microscope | Zeiss | ||
Leptomycin B | Sigma | L2913 | |
PBS | Biological Industries | 02-023-1A | |
Homogenizer motor/control/chuck/90-degree clamp | Glas-Col | 099C K5424CE | |
MicroCL 17R Centrifuge, Refrigerated | Thermo Scientific | 75002455 |
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Source: Shani, T., et al. Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells. J. Vis. Exp. (2017)
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