Tris buffered saline tbs

Human and cynomolgus monkey tissue and hiPSC/SEAM-derived cell sheets were embedded in Optimal Cutting Temperature Compound (Sakura Fineteck, Tokyo, Japan) and sectioned at 10 μm thickness. The sections were fixed in 2–4% paraformaldehyde (Wako), washed three times in Tris-buffered saline (TBS) (TaKaRa Bio, Shiga, Japan) for 5 min, and incubated for 1 h in TBS containing 5% normal donkey serum (Jackson immunoReseach, West Grove, PA, USA) and 0.3% Triton X-100 (Sigma-Aldrich, St. Louis, MO, USA) to block non-specific reactions. Next, cells/tissues were incubated with the primary antibodies listed in the key resources table at 4°C overnight and stained with Hoechst 33342 (Wako) and Alexa Fluor-conjugated secondary antibodies (Life Technologies) for 1 h at room temperature. Stained cells or tissues were observed using an Axio Imager D1 (Carl Zeiss).

Phosphate-buffered saline (PBS) was purchased from Gibco/Life technologies (Carlsbad, CA). Tris-buffered saline (TBS) tablets were purchased from Takara Bio (Otsu, Japan). Urea, Tween 20, and glucose were purchased from Sigma-Aldrich (St Louis, MO). Sodium dodecyl sulfate (SDS) was purchased from WAKO (Tokyo, Japan).