Claudin 1 Polyclonal Antibody

Protocol tips

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	-Rabbit (1:200)

Protocol tips
-Rabbit (1:200)

Publication protocol

Ileum segments were placed in RIPA buffer (20 mmol/L Tris, pH 7.4, 1% Triton X-100, 1% deoxycholate, 0.1% SDS, 100 mmol/L NaCl) containing protease inhibitors (Complete Mini, cat. 11374600; Roche, Basel, Switzerland) and the phosphatase inhibitors calyculin A and sodium orthovanadate (Sigma, St. Louis, MO) and homogenized. Debris was removed by centrifugation and protein concentration was determined using the Pierce 660-nm Protein Assay Reagent (cat. 1861426; Thermo Scientific). For immunoblotting, 15–20 mg of protein was separated on a SDS–polyacrylamide gel electrophoresis, transferred to nitrocellulose, blocked in tris-buffered saline with 0.1% tween-20 (TBST) þ 5% milk, and incubated in primary antibody overnight at 4C (Table 1). Blots were washed in TBST at room temperature and incubated with secondary antibody followed by imaging using the LICOR-Odyssey system.

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Manufacturer protocol

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