NOD2 Antibody (2D9): sc-56168

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:600) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Protocol tips
-Mouse (1:600) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Publication protocol

Tissue sections were fixed using 4% paraformaldehyde (PFA) for 60 minutes at 4°C followed by 3 wash steps in phosphate buffered saline (PBS) of 5 minutes each. Blocking was performed in 5% goat serum for 30 minutes followed by incubation with primary antibodies for 2 hours at room temperature at the following dilutions: rabbit anti-NOD1 1 : 500 (Sigma-Aldrich) and rabbit anti-NOD2 1 : 500 (Sigma-Aldrich). Secondly, fluorochrome-conjugated antibodies were diluted 1 : 300 (Alexa 555) and slices were incubated for 1 hour at room temperature within this solution. Nuclear counterstaining was performed using SYTOX green, 1 : 20000 (Molecular Probes) for 30 minutes at room temperature. The stained sections were mounted with Mowiol (Carl Roth) and dried over night at 4°C. Fluorescence imaging was performed using a confocal microscope (LSM 510, Carl Zeiss, San Diego, CA, USA, and DM IRB, Leica Microsystems, Inc., Buffalo Grove, IL, USA).

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Manufacturer protocol

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