Anti-AKT (phospho T308) antibody (ab38449)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.	-Rabbit (1:1000) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Upstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.
Protocol tips
-Rabbit (1:1000) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Publication protocol

Transfected OSCC cells were lysed using RIPA buffer (Thermo Scientific, Belmont, MA, U.S.A.) at 4°C for 30 min. Proteins in OSCC cells were extracted and separated through 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and transferred onto a PVDF membrane (Roche) via electroblotting. The membranes were blocked in 5% nonfat milk for 1 h at room temperature and then incubated at 4°C overnight with primary antibodies, including anti-PI3K (ab32089), anti-p-PI3K (ab182651), anti-Akt (ab179463), anti-p-Akt (ab38449), anti-mTOR (ab2732), anti-p-mTOR (ab109268), and anti-GAPDH (ab181602) antibodies. Next, the membrane was incubated for 1.5 h at room temperature using second antibody (1:2000, Abcam, China). Target band of protein was visualized using ECL Western blotting kit (Millipore, Boston, MA, U.S.A.).

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Manufacturer protocol

Download the product protocol from Abcam for Anti-AKT (phospho T308) antibody (ab38449) below.

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