Atg12 Antibody (Mouse Specific) #2011

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Dilute primary Ab at 1:100 and incubate overnight at 4 °C and 1 h at room temperature

Protocol tips
Dilute primary Ab at 1:100 and incubate overnight at 4 °C and 1 h at room temperature

Publication protocol

Cells were grown on 3 cm Petri dish and were fixed in 4% paraformaldehyde for 15 min. Cells were washed two times in phosphate-buffered saline (PBS), permeabilized with 0.1% Triton X-100 for 15 min, followed by another wash in PBS and block in PBS containing 5% bovine serum albumin (BSA) for 1 h. Primary and secondary antibodies in PBS containing 5% BSA were applied overnight at 4 °C and 1 h at room temperature, respectively. Nuclei were costained with 1 μg/ml Hoechst 33342. Fluorescence images were taken using a Nikon Eclipse TE200 epifluorescence microscope with NIS-Elements AR3.2 software or EVOS FL Auto (Life Technologies, Bothell, WA, USA). For manual quantification of the puncta formation at least three optical fields with at least 50 cells per experimental condition were analyzed. Data from repeated experiments were subjected to statistical analysis.

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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for Atg12 Antibody (Mouse Specific) #2011 below.

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