Laemmli Lysis-buffer

Protocol tips

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	Spore samples were harvested at 72 h, resuspended to 1 × 107 spores/100 μl in 1× Laemmli loading buffer, and heated to 95°C for 5 min. Spore extracts were separated on 3 to 8% NuPAGE Novex Tris-acetate minigels and blotted onto polyvinylidene difluoride (PVDF).

Protocol tips
Spore samples were harvested at 72 h, resuspended to 1 × 107 spores/100 μl in 1× Laemmli loading buffer, and heated to 95°C for 5 min. Spore extracts were separated on 3 to 8% NuPAGE Novex Tris-acetate minigels and blotted onto polyvinylidene difluoride (PVDF).

Publication protocol

Spore samples were harvested at 72 h, resuspended to 1 × 107 spores/100 μl in 1× Laemmli loading buffer, and heated to 95°C for 5 min. Spore extracts were separated on 3 to 8% NuPAGE Novex Tris-acetate minigels and blotted onto polyvinylidene difluoride (PVDF). The membrane was probed with a 1:5,000 dilution of anti-β-O-GlcNAc (Covance, Montreal, Canada) in PBS–0.1% Tween 20 (PBS-T). Reactivity was detected with anti-mouse IgM horseradish peroxidase (HRP) conjugate (Sigma-Aldrich, Oakville, Canada) secondary antibody at a 1:10,000 dilution in PBS-T. Blots were imaged with an ECL Prime Western blotting detection kit (GE Healthcare, Baie D'Urfe, QC, Canada) according to the manufacturer's instructions, followed by exposure to X-ray film.

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