Protein Localization in the Cyanobacterium Anabaena sp. PCC7120 Using Immunofluorescence Labeling
- Resource Type
- Authors
- Carla Trigo; Mónica Vásquez; Derly Andrade
- Source
- Bio Protoc
- Subject
- chemistry.chemical_classification
Fluorophore
medicine.diagnostic_test
biology
Strategy and Management
Mechanical Engineering
Metals and Alloys
Peptide
Immunofluorescence
Primary and secondary antibodies
Protein subcellular localization prediction
Industrial and Manufacturing Engineering
Autofluorescence
chemistry.chemical_compound
chemistry
Biochemistry
Fluorescence microscope
medicine
biology.protein
Methods Article
Antibody
- Language
- English
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.