LC3B Antibody Kit for Autophagy

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	According to manufacturer's protocol, i.e; 0.5 microgram/ml antibody in blocking buffer

Protocol tips
According to manufacturer's protocol, i.e; 0.5 microgram/ml antibody in blocking buffer

Publication protocol

Fluorescence images
MCs were stained with 1 µg/ml of propidium iodide (PI, Sigma) on ice, then attached to a glass slide by using Shandon Cytospin 2 cytocentrifuge (Thermo Fisher Scientific). Prior to Cytospin Cells were resuspended at 106/ml. Each slide had the same amount of spun cells. Slides were mounted in ProLong Anti-Fade reagent with DAPI (Molecular Probes). For autophagy assay, we used LC3B antibody Kit for autophagy (Invitrogen) according to the manufacturers' instructions. We imaged the MCs using the Bx51 research microscope (Olympus) and X-Cite 120 fluorescence illumination systems (EXFO Photonic Solutions).

Autophagy assay for mast cell
We used LC3B Antibody Kit for Autophagy (Invitrogen) according to manufacturer's instructions.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for LC3B Antibody Kit for Autophagy below.

Download manufacturer protocol

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