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. 2010 Oct 15;285(51):40097–40103. doi: 10.1074/jbc.M110.144626

FIGURE 2.

FIGURE 2.

Glycan analysis of brain endothelial APP. A, BMEC lysates (20 μg of protein) were incubated in the presence or absence of PNGase and with or without sialidase. The digested samples were analyzed using the anti-APP C15 antibody. B, BMEC lysates (6 μg of protein) were incubated in the presence or absence of sialidase O-glycosidase and then immunostained with the anti-APP C15 antibody. C, APP precipitated from BMEC lysates (0.25 mg of protein) using the anti-APP C15 antibody (5 μg) was incubated in the presence or absence of sialidase (4 milliunits). The digested samples were subjected to Western blot analysis with the anti-APP 22C11 antibody or PNA lectin blot analysis. D, BMEC lysates (100 μg of protein) were incubated with Sambucus sieboldiana agglutinin (SSA)-, Maackia amurensis agglutinin (MAA)-, E4-phytohemagglutinin (PHA-E4)-, concanavalin A (ConA)-, Ricinus communis agglutinin (RCA)120-, or jacalin-agarose for 16 h at 4 °C. The lectin-precipitated samples were washed and analyzed by Western blotting with the anti-APP C15 antibody. IB, immunoblot.