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Updated: Oct 8, 2026
Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass
Published on: August 10, 2017
Protocol for confocal microscopy-based localization of chorismate mutase protein of Mycobacterium tuberculosis in
Vandana Maurya1, Sangita Mukhopadhyay2
1Laboratory of Molecular Cell Biology, BRIC-Centre for DNA Fingerprinting and Diagnostics (CDFD), Inner Ring Road, Uppal, Hyderabad 500039, Telangana, India; Graduate Studies, Regional Centre for Biotechnology, Faridabad 121001, Haryana (NCR Delhi), India.
Abstract:
Here, we present a confocal microscopy protocol to determine the localization of Mycobacterium tuberculosis chorismate mutase (MtbCM) in trans-Golgi compartment. We co-transfect HEK293 cells with either pEGFP-C1-MtbCM and mCherry-TGOLN2 (trans-Golgi marker) or pEGFP-C1 and mCherry-TGOLN2 as a control, using Lipofectamine 2000. We describe steps for constructing pEGFP-C1-MtbCM and performing co-transfection and image acquisition using confocal microscopy. This protocol provides a representative procedure to determine the subcellular localization of the mycobacterial protein to the trans-Golgi compartment. For complete details on the use and execution of this protocol, please refer to Pal et al.1.
