Abstract:
:Multicolor immunostaining analysis is often a desirable tool in cell biology for most researchers. Nonetheless, this is not an easy task and often not affordable by many laboratories as it might require expensive instrumentation and sophisticated analysis software. Here, we describe a simple protocol for performing sequential immunostainings on two different Drosophila specimens. Our strategy relies on an efficient and reproducible method for removal primary antibodies and/or fluorophore-conjugated secondary antibodies that does not affect antigene integrity. We show that alternation of multiple rounds of antibody incubation and removal on the same slide, followed by registration of the same DAPI-stained image, provides a simple framework for the sequential detection of several antigens in the same cell. Given that the sample fixation procedures used for Drosophila tissues are compatible with most specimen processing protocols, we can envisage that the multicolor immunostaining strategy presented here can be also adapted to different samples including mammalian tissues and/or cells.
journal_name
J Cell Physioljournal_title
Journal of cellular physiologyauthors
Cipressa F,Di Giorgio ML,Cenci Gdoi
10.1002/jcp.24506subject
Has Abstractpub_date
2014-06-01 00:00:00pages
683-7issue
6eissn
0021-9541issn
1097-4652journal_volume
229pub_type
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