Typical cross-contamination between cytosolic and nuclear fractions is about 10%. The method recovers primarily soluble proteins from the nucleus, not so much proteins bound in chromatin or integral membrane proteins of the nuclear envelope.

2021年6月18日 · An important advantage of this protocol is in its adaptation for simple and reliable separation of nuclear and cytoplasmic fractions, which allows one to study the distribution of endogenous and exogenous shuttle proteins in the Xenopus embryo cells in response to various experimental stimuli.

2011年12月13日 · We present a simple method, based on partial membrane permeabilization with detergent followed by density gradient centrifugation, that provides rapid separation of cytosolic, nucleosolic, nuclear insoluble, and membrane components in …

Here, we compared different nucleus/cytoplasm fractionation methods and selected the best-suited approach for the separation of nuclear and cytoplasmic contents. The described methodology is based on stepwise lysis of cells and washing of the resulting nuclei using non-ionic detergents, such as NP-40.

2024年7月23日 · Nuclear extraction enables you to separate nuclear and cytoplasmic cellular components and associated samples. Commercial kits simplify the process but seldom explain what the chemicals do, making troubleshooting difficult when it fails.

Thermo Scientific Subcellular Protein Fractionation Kit enables stepwise separation and extraction of cytoplasmic, membrane, nuclear-soluble, chromatin-bound and cytoskeletal proteins from mammalian cultured cells or tissue.

2011年12月13日 · We present a simple method, based on partial membrane permeabilization with detergent followed by density gradient centrifugation, that provides rapid separation of cytosolic, nucleosolic, nuclear insoluble, and membrane components in …

2024年5月20日 · Biochemical fractionation is a technique used to isolate and separate distinct cellular compartments, critical for dissecting cellular mechanisms and molecular pathways. Herein we outline a biochemical fraction methodology for isolation of ultra-pure nuclei and cytoplasm.

2010年11月10日 · Separation of nuclear and cytoplasmic proteins by differential centrifugation in non-ionic detergent. Panel A, B, C and D show fractionation results from HeLa, HCT116, HEK293 and Hs68 cells, respectively.

Remove the cytoplasmic extract from the pellet to a clean tube. Wash the nuclei with 100 μl of CE buffer without detergent. Be careful to resuspend the fragile nuclei gently. Add 1 pellet volume NE buffer to nuclear pellet (approximately 50 μl). Adjust the salt concentration to 400 mM using 5 M NaCl (add ~35 μl).