Journal: Heliyon

Article Title: Immunoproteomic approach identifies a putative virulence chaperone DnaK protein as a candidate diagnostic marker and therapeutic target for Pythium insidiosum infection

doi: 10.1016/j.heliyon.2025.e42487

Figure Lengend Snippet: SDS-PAGE and Western blot analysis of P. insidiosum 's synthesized proteins. Nineteen coding sequences of the recruited target proteins S01-S21 (except S03 and S04) serve as a protein synthesis template in the CFPS reaction, and the resulting products are checked for their presence using SDS-PAGE stained with Coomassie blue dye ( A ) and their immunoreactivity using Western blot analysis probed against the mouse anti-6x-His tag antibody ( B ). Green fluorescent protein (GFP) is a positive control for the CFPS reaction. An arrowhead indicates a target synthesized protein. The molecular weight markers (M) are shown on the left of each figure. The original unprocessed images of are available as supplementary files: Original_Fig2A_S01-02_S11-S14_GFP.tif, Original_Fig2A_S05-06.tif, Original_Fig2A_S07-10.tif, Original_Fig2A_S015-21.tif, Original_Fig2B_S01-S02_S11-S14_GFP.tif, Original_Fig2B_S05-06.tif, Original_Fig2B_S07-10.tif, and Original_Fig2B_S015-21.tif.

Article Snippet: Up to 16 different proteins can be synthesized simultaneously using a second-round PCR product as a template for protein synthesis and a commercial CFPS kit from Bioneer with an automated protein synthesis machine from the same company.

Techniques: SDS Page, Western Blot, Synthesized, Staining, Positive Control, Molecular Weight