α Tubulin Antibody (TU-02): sc-8035

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:1000) -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 (1:1000) -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

Dosal epidermal tissues were treated with dispase for overnight at 4 °C. The epidermis was separated and lysed in protein extraction solution (Intron) after homogenization using tissue lyser (QIAGEN). Epidermal extracts were centrifuged for 15 min at 15,000 rpm. Total protein was measured using a BCA protein assay kit (Thermo Scientific, Rockford, IL). Samples (20–30 μg protein per lane) were run on SDS-polyacrylamide gels, transferred onto nitrocellulose membranes and incubated with appropriate antibodies (1:1,000) for overnight at 4 °C with gentle agitation. Blots were then incubated with peroxidase-conjugated secondary antibodies (1:2,000) for 30 minutes at room temperature, and visualized by enhanced chemiluminescence (TransLab). The following primary antibodies were used in this study: Crif1 (Santacruz, sc-134882), β-catenin (Santacruz, sc-7199), Actin (Santacruz, sc-1615), LaminB (Santacruz, sc-6216), α-tubulin (Santacruz, sc-8035), MTCO1 (Abcam, ab14705), Involucrin (Abcam, ab28057), Filaggrin (Abcam, ab81468), and Loricrin (Corvance, PRB-145P).

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