Imaginal Disc Immunostaining Protocol

Immuno-staining of imaginal discs was according to standard protocols using Hairless anti-A (1:500, from guinea pig) [21 (link)], mouse anti-Cut (1:25) or anti-Wingless (1:25) (developed by G. Rubin and S.M. Cohen, respectively; obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Dept of Biology, Iowa City, IA 52242) and rabbit anti-GFP (1:200, Santa Cruz Biotech, Dallas, USA). Goat secondary antibodies coupled to FITC, Cy3 or Cy5 were from Jackson Immuno-Research (Dianova, Hamburg, Germany). Tissue was mounted in Vectashield (Vector Labs, Eching, Germany), and examined using a BioRad MRC1024 confocal microscope and LaserSharp 2000TM software (Carl Zeiss, Jena, Germany).

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Praxenthaler H., Smylla T.K., Nagel A.C., Preiss A, & Maier D. (2015). Generation of New Hairless Alleles by Genomic Engineering at the Hairless Locus in Drosophila melanogaster. PLoS ONE, 10(10), e0140007.

Publication 2015

rabbit anti-GFP FITC Cy3 Vectashield MRC1024 confocal microscope LaserSharp 2000TM software

Antibodies Confocal microscope Fitc Goat Guinea pig Hybridoma Imaginal discs Mouse Rabbit Tissue Vector

Corresponding Organization : University of Hohenheim

Top 5 similar protocols

Variable analysis

independent variables

Hairless anti-A (1:500, from guinea pig)
Mouse anti-Cut (1:25)
Anti-Wingless (1:25)

dependent variables

Hairless anti-A expression
Cut expression
Wingless expression

control variables

Tissue mounting in Vectashield
Imaging using a BioRad MRC1024 confocal microscope and LaserSharp 2000TM software

positive controls

Rabbit anti-GFP (1:200, Santa Cruz Biotech, Dallas, USA)

negative controls

Not explicitly mentioned

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