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Mini protean tgx precase gel

Manufactured by Bio-Rad

The Mini-Protean TGX Precast Gel is an electrophoresis gel designed for protein separation and analysis. It is a ready-to-use, pre-cast polyacrylamide gel that provides consistent and reliable results.

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2 protocols using mini protean tgx precase gel

1

Gel Electrophoresis Protein Separation

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Proteins were separated on a 12% Mini-Protean TGX Precase Gel (BioRad) before being incubated overnight in GelCode (Thermo Fisher). Gels were washed with deionized water and imaged using GeneSys.
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2

Quantifying Shigella Secreted Proteins

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Proteins were separated on a 12% Mini-Protean TGX Precase Gel (BioRad) before being transferred to polyvinylidene difluoride (PVDF) membrane. The membrane was blocked with 10% (wt/vol) non-fat milk buffer in 1× DPBS and then incubated with anti-IpaB, anti-IpaC, and anti-IpaD antibody (1:10,000) (kindly donated by the Wendy Picking Laboratory) diluted in 10% (wt/vol) non-fat milk buffer overnight at 4°C on an orbital shaker. Following washing, the membrane was incubated with secondary goat anti-rabbit 680 nm (Thermo Fisher), and proteins were visualized using 700-nm wavelength setting on a LI-COR Odyssey Laser Scanner. Whole cell lysates were normalized using DnaK (Invitrogen) as a loading control, and densitometry (ImageJ) (66 (link)) was utilized to compare loaded protein levels. There is no other defined protein in Shigella that is secreted in a consistent quantity for use as a control. Densitometry (ImageJ) for secreted fractions were determined by assuming 2457T without bile salts was 1 and comparing all secreted levels protein levels to it.
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