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Fractionation and Extraction of Crude Nuclear Proteins From Arabidopsis Seedlings

Authors: Jiajia Zhao, Jinsong Bao and Feifei Xu, date: 01/20/2022, view: 1115, Q&A: 0

[Abstract]

The plant nucleus is an important subcellular organelle that contains the genome, ribosomal RNA, and regulatory proteins, and performs a central role in the functioning and metabolism of the cell. Fractionation of intact nuclei is a crucial process to elucidate the function of nuclear proteins. Here, we present a simple method for the

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ChIP-Seq from Limited Starting Material of K562 Cells and Drosophila Neuroblasts Using Tagmentation Assisted Fragmentation Approach

Authors: Junaid Akhtar, Piyush More and Steffen Albrecht, date: 02/20/2020, view: 2773, Q&A: 0

[Abstract] Chromatin immunoprecipitation is extensively used to investigate the epigenetic profile and transcription factor binding sites in the genome. However, when the starting material is limited, the conventional ChIP-Seq approach cannot be implemented. This protocol describes a method that can be used to generate the chromatin profiles from as low as ...

NP-40 Fractionation and Nucleic Acid Extraction in Mammalian Cells

Authors: Alvaro E. Galvis, Hugh E. Fisher and David Camerini, date: 10/20/2017, view: 12205, Q&A: 4

[Abstract] This technique allows for efficient, highly purified cytoplasmic and nuclear-associated compartment fractionation utilizing NP-40 detergent in mammalian cells. The nuclear membrane is not disturbed during the fractionation thus leaving all nuclear and perinuclear associated components in the nuclear fraction. This protocol has been modified from ...

Nuclei Isolation from Nematode Ascaris

Authors: Yuanyuan Kang, Jianbin Wang and Richard E. Davis, date: 05/05/2017, view: 7967, Q&A: 0

[Abstract] Preparing nuclei is necessary in a variety of experimental paradigms to study nuclear processes. In this protocol, we describe a method for rapid preparation of large number of relatively pure nuclei from Ascaris embryos or tissues that are ready to be used for further experiments such as chromatin isolation and ChIP-seq, nuclear RNA ...

Differential Salt Fractionation of Nuclei to Analyze Chromatin-associated Proteins from Cultured Mammalian Cells

Authors: Christin Herrmann, Daphne C. Avgousti and Matthew D. Weitzman, date: 03/20/2017, view: 13668, Q&A: 0

[Abstract] Nucleosomes are the core units of cellular chromatin and are comprised of 147 base pairs (bp) of DNA wrapped around an octamer of histone proteins. Proteins such as chromatin remodelers, transcription factors, and DNA repair proteins interact dynamically with chromatin to regulate access to DNA, control gene transcription, and maintain genome ...

An Efficient Procedure for Protoplast Isolation from Mesophyll Cells and Nuclear Fractionation in Rice

Authors: Mehdi Jabnoune, David Secco, Cécile Lecampion, Christophe Robaglia, Qingyao Shu and Yves Poirier, date: 03/05/2015, view: 21529, Q&A: 1

[Abstract] Plant protoplasts, a proven physiological and versatile cell system, are widely used in high-throughput analysis and functional characterization of genes. Green protoplasts have been successfully used in investigations of plant signal transduction pathways related to hormones, metabolites and environmental challenges. This protocol, adapted from ...

Protocol for Preparation of Nuclear Protein from Mouse Lungs

Authors: Zuoxiang Xiao and Qun Jiang, date: 06/05/2014, view: 12855, Q&A: 0

[Abstract] This protocol describes how to extract nuclear protein from mouse lungs, including tissue preparation, stepwise lysis of cells and centrifugal isolation of nuclear protein fraction. This is an efficient method to get comparable nuclear protein extracts from lung tissues.

Cell Cycle Analysis in the Vertebrate Brain Using Immunolabeled Fresh Cell Nuclei

Authors: Noelia Lopez-Sanchez and Jose M. Frade, date: 11/20/2013, view: 15768, Q&A: 0

[Abstract] Flow cytometry, a standard technique used for quantitative analysis of isolated cells, is routinely employed by immunologists and oncologists to study DNA content, protein expression, and other functional parameters in blood and tumor cells. Unfortunately, the use of this technique by neurobiologists has been hampered by the complexity of the ...

Micrococcal Nuclease (MNase) Assay of Arabidopsis thaliana Nuclei

Authors: Laia Armengot and Jordi Moreno-Romero, date: 04/05/2013, view: 22097, Q&A: 3

[Abstract] Micrococcal nuclease (MNase) is able to produce double-strand breaks within nucleosome linker regions. The efficiency of MNase digestion depends on the degree of chromatin compaction, being more easily digested the regions of less compacted chromatin. The MNase protocol described here can be used to asses changes in the chromatin structure of ...

Nuclear Extraction from Arabidopsis thaliana

Authors: Fang Xu and Charles Copeland, date: 12/20/2012, view: 31473, Q&A: 14

[Abstract] This protocol is to isolate nuclei from Arabidopsis cells. They can be further used for other experiments, such as nuclear protein detection, nuclear protein immunoprecipitation and so on.

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