Nanog Antibody (J-29): sc-81961

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

Western blot was performed following the standard protocol [22, 31]. The cell lysates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and were transferred to the polyvinylidene fluoride membranes. After the membranes were blocked, the diluted primary antibodies, MDR1 (MAB4163, 1:1000, Chemicon, Temecula, CA, USA ), ABCG2 (MAB4155, 1:1000, Millipore, Billerica, MA, USA), α-Tubulin (sc-5286, 1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), E-cadherin (sc-7870, 1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), Snail (sc-28199, 1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), Nanog (sc-81961, 1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), Oct4 (GTX101497, 1:1000, Gene Tex, San Antonio, TX, USA), in Tris-buffered saline with Tween (TBST) buffer containing 3% non-fat milk were added to the membranes and incubated at 4°C overnight. On the second day, the goat anti-mouse conjugated secondary antibody was added at room temperature for 1 h. The immunoblots were developed using an enhanced chemiluminescence system and visualized on X-ray film.

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