K-Ras Antibody (F234): sc-30

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:200) -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 (1:200) -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

For immunoblot, cell pellets and tissues were lysed with SDS-PAGE lysis buffer, boiled and resolved with SDS-PAGE electrophoresis. Proteins were then transferred to nitrocellulose membranes, which were blocked with 5% non-fat milk and probed with corresponding antibodies including mouse anti-KRAS monoclonal antibody (sc-30, 1:200, Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-ME1 monoclonal antibody (sc-100569, 1:1000, Santa Cruz Biotechnology), rabbit anti-P65 polyclonal antibody (A2547, 1:1000, Abclonal), mouse anti-Flag monoclonal antibody (F3165, 1:10 000, Sigma-Aldrich) and mouse anti-β-actin monoclonal antibody (sc-47778, 1:2000, Santa Cruz Biotechnology). The blots were then incubated with a 1:1000–1:5000 dilution of HRP-conjugated IgG (Santa Cruz Biotechnology), washed and subjected to chemiluminescent detection as previously described.

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