Bacterial Culture Filtrate Protein Concentration using an Ultrafiltration Device

Published: November 30, 2023

Abstract

Source: Ryan, J. M., et al. Mycobacterium tuberculosis Extracellular Vesicle Enrichment through Size Exclusion Chromatography. J. Vis. Exp. (183) (2022)

This video demonstrates the isolation and concentration of bacterial culture filtrate proteins, which include bacterial extracellular vesicles, using ultrafiltration. The concentrated extracellular vesicles serve as diagnostic markers for disease detection.

Protocol

1. Preparation of crude Mycobacterium tuberculosis (Mtb) extracellular vesicle (EV) concentrate NOTE: For detailed procedures on the cultivation of Mtb and preparation of culture filtrate protein (CFP). It is recommended that bacterial culture media is free from growth supplements with EV-containing or proteinaceous components, such as Oleic Albumin Dextrose Catalase (OADC), and detergents such as Tween. It is also recommended that the bacterial culture…

Divulgaciones

The authors have nothing to disclose.

Materials

Automatic Fraction Collector IZON Science AFC-V1-USD
Centricon Plus – 70 Centrifugal filter, 100 kDa cutoff Millipore Sigma UFC710008 Ultrafiltration device used in step 1.1
Phosphate-buffered Saline, 1X without calcium and magnesium Corning 21-040-CV
Nitrocellulose membrane, Roll, 0.2 μm BioRad 1620112

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Citar este artículo
Bacterial Culture Filtrate Protein Concentration using an Ultrafiltration Device. J. Vis. Exp. (Pending Publication), e21874, doi: (2023).

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