A simple approach for multicolor immunofluorescence staining in different Drosophila cell types.

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 Physiol

authors

Cipressa F,Di Giorgio ML,Cenci G

doi

10.1002/jcp.24506

subject

Has Abstract

pub_date

2014-06-01 00:00:00

pages

683-7

issue

6

eissn

0021-9541

issn

1097-4652

journal_volume

229

pub_type

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