Campus Units

Biochemistry, Biophysics and Molecular Biology, Roy J. Carver Department of

Document Type

Article

Publication Version

Accepted Manuscript

Publication Date

8-2009

Journal or Book Title

Methods

Volume

48

Issue

43

First Page

387

Last Page

397

DOI

10.1016/j.ymeth.2009.02.019

Abstract

The giant polytene chromosomes from Drosophila third instar larval salivary glands provide an important model system for studying the architectural changes in chromatin morphology associated with the process of transcription initiation and elongation. Especially, analysis of the heat shock response has proved useful in correlating chromatin structure remodeling with transcriptional activity. An important tool for such studies is the labeling of polytene chromosome squash preparations with antibodies to the enzymes, transcription factors, or histone modifications of interest. However, in any immunohistochemical experiment there will be advantages and disadvantages to different methods of fixation and sample preparation, the relative merits of which must be balanced. Here we provide detailed protocols for polytene chromosome squash preparation and discuss their relative pros and cons in terms of suitability for reliable antibody labeling and preservation of high resolution chromatin structure.

Comments

This is a manuscript of an article published as Johansen, Kristen M., Weili Cai, Huai Deng, Xiaomin Bao, Weiguo Zhang, Jack Girton, and Jørgen Johansen. "Polytene chromosome squash methods for studying transcription and epigenetic chromatin modification in Drosophila using antibodies." Methods 48, no. 4 (2009): 387-397. doi: 10.1016/j.ymeth.2009.02.019. Posted with permission.

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Copyright Owner

Elsevier Inc.

Language

en

File Format

application/pdf

Published Version

Share

COinS