Preparation of Subcellular Fractions from Leukemic Cells: A Method to Generate Cellular and Nuclear Fractions from Leukemic Cells
Published: April 30, 2023
Abstract
Source: Hay, J., et al. Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking. J. Vis. Exp. (2019).
This video describes the preparation of subcellular fractions from leukemic cells which is a method to generate cellular and nuclear fractions from leukemic cells. The nuclear and cytoplasmic fractions can be used for further downstream analysis.
Protocol
1. Preparation of Subcellular Fractions from CLL Cells NOTE: When planning the experimental set-up, include a well of unstimulated/untreated cells from which the whole cell extract can be generated. Perform the desired stimulation and/or drug treatment of the MEC1 CLL cell line or isolated primary CLL cells using 10 – 20 x 106 cells/condition. Cells will then be used for subcellular fractionation or to generate whole cell extract as expla…
Declarações
The authors have nothing to disclose.
Materials
| 1.5 mL microcentrifuge Tubes | Griener Bio one | 616201 | |
| EDTA | Sigma | EDS | |
| Labnet VX100 | Fisher Scientific | Vortex | |
| Nucelar Extract Kit | Active Motif | 40010 | |
| PBS Tablets | Fisher Scientific | BR0014G | |
| Sigma 3-16P | SciQuip | Centrifuge | |
| Sigma 1-15PK | SciQuip | Centrifuge |
Citar este artigo
Preparation of Subcellular Fractions from Leukemic Cells: A Method to Generate Cellular and Nuclear Fractions from Leukemic Cells. J. Vis. Exp. (Pending Publication), e20266, doi: (2023).