c-Myc Antibody (9E10): sc-40

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:500) -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 (1:500) -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 Western blot, protein samples were extracted from transfected S2 cells and analyzed by standard biochemical procedures. Primary antibodies used include: anti-α-tubulin (mouse, Sigma B5–1–2, 1:500000), anti-p-Ser-PKC (rabbit, Cell Signaling #2261 S, 1:300), anti-Flag-HRP (rabbit, Sigma A8592, 1:1000), anti-HA-HRP (rat, Roche 3F10, 1:1000), or anti-c-Myc-HRP (mouse, Santa Cruz Biotechnology sc-40 HRP, 1:500). For Co-IP analysis, protein lysates from S2 cells were harvested and lysed using the standard protocols. Flag antibody-conjugated beads (mouse, sigma A2220) or the anti-HA affinity matrix (rat, Roche 11815016001) were used to pull down the corresponding proteins for 2–2.5 hours at 4 °C. After serial washes with lysis buffer, resins were subjected to SDS-PAGE for analysis with antibodies described above. For experiments with drug treatments, drugs used were: MG132 (50 μM, BioVision), E64 (1 μM, Merck Millipore), OA (0.1 μM, Sigma 75320), and PMA (0.01 μM, Sigma P8139).

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