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Total RNA for Northern blots was prepared using Qiagen RNeasy Midi kits.
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In complementary experiments, Western blots were prepared using control tissue obtained from an adult rat-brain microsomal preparation or the total lysate from the SVZ eNPC (Figure 2 C).
Blots were prepared using antibodies as described [23].
Lysates for western blots were prepared using the PARIS™ kit with a protease inhibitor cocktail (Sigma).
Colony blots were prepared using Whatmans 541 filter paper (Whatman, England).
Western blots were prepared using extracts of H. polymorpha and P. chrysogenum cells, obtained using the TCA method [27], and decorated using antibodies raised against IPNS, ACVS, IAT, PCL [8], [28], or the His6 tag (Santa Cruz Biotechnology, INC).
After electro-transfer to a PVDF membrane (pore diameter, 0.45mM), western blots were prepared using antibody 369 (which recognizes the C-terminal region of APP and APP-CTFs) or anti-b-actin (Santa Cruz Biotechnology, sc-4778).
Western blots were prepared using standard protocols.
Northern blots were prepared using the NorthernMax-Gly kit (Ambion) following the manufacturer's instructions.
RNA blots were prepared using 10 15 μg of total RNA isolated from leaf tissue using the TRIzol protocol (Invitrogen).
Western blots were prepared using crude extracts of TCA precipitated cells and decorated with a polyclonal α-IPNS antiserum.
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