NDLI: Reliable detection of epigenetic histone marks and nuclear proteins in tissue cryosections

Content Provider	Springer Nature Link
Author	Eberhart, Anja Kimura, Hiroshi Leonhardt, Heinrich Joffe, Boris Solovei, Irina
Copyright Year	2012
Abstract	Nuclear processes in real tissues often are significantly different from those in cultured cells. However, immunostaining on tissue sections needs long fixation which masks antigens and, respectively, antigen retrieval which restores antigen accessibility. These treatments affect the immunostaining results and complicate their interpretation. The problem is especially significant for nuclear antigens which often are very sensitive to both fixation and antigen retrieval. We targeted this problem by a study of several histone modifications and nuclear proteins in tissue sections of mouse retina which contains cells with both conventional and unique inverted nuclei. In the latter, the main chromatin classes form separate concentric shells which simplifies evaluation of the signal distribution. We show that as a rule, longer fixation demands longer antigen retrieval time. Nevertheless, antigens are remarkably diverse in this respect and need individual adjustment. We suggest a robust procedure for immunostaining on sections, that is, a method that allows controlling the differences in immunostaining caused by differences in fixation time and antigen retrieval duration, so that immunostaining protocol can be quickly optimized.
Starting Page	849
Ending Page	858
Page Count	10
File Format	PDF
ISSN	09673849
Journal	Chromosome Research
Volume Number	20
Issue Number	7
e-ISSN	15736849
Language	English
Publisher	Springer Netherlands
Publisher Date	2012-11-02
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Nuclear architecture epigenetic histone marks nuclear proteins mouse tissues mouse retina cryosections immunostaining antigen retrieval tissue fixation Cell Biology Human Genetics Animal Genetics and Genomics Plant Genetics & Genomics
Content Type	Text
Resource Type	Article
Subject	Genetics

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