Bio-Protocol (Jun 2017)

Protein Localization in the Cyanobacterium Anabaena sp. PCC7120 Using Immunofluorescence Labeling

  • Carla Trigo,
  • Derly Andrade,
  • Mónica Vásquez

DOI
https://doi.org/10.21769/BioProtoc.2318
Journal volume & issue
Vol. 7, no. 11

Abstract

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Techniques such as immunoflorescence are widely used to determine subcellular distribution of proteins. Here we report on a method to immunolocalize proteins in Anabaena sp. PCC7120 with fluorophore-conjugated antibodies by fluorescence microscopy. This method improves the permeabilization of cyanobacterial cells and minimizes the background fluorescence for non-specific attachments. In this protocol, rabbit antibodies were raised against the synthetic peptide of CyDiv protein (Mandakovic et al., 2016). The secondary antibody conjugated to the fluorophore Alexa488 was used due to its different emission range in comparison to the autofluorescence of the cyanobacterium.