Lysates from transfected 293T cells or human tissue from frontal lobe cortex were probed in Western blot by the following antibodies: Anti-FLAG (M2),Sigma; anti-HA (3F10),Roche; anti-Ubiquitin (1B3), MBL; anti-GluR4 (AB1508),Chemicon; anti-NSE (BBS/NC/V1-H14) Dako; Anti-myc (9E10), anti-HA (Y-11), anti-Ubiquitin (FL-76), and anti-actin (2) all from Santa Cruz. Anti-UCH-L1 (gift of K. Wade, Japan) was also used in Western Blot assays.
Immunohistochemistry on mouse brains employed Anti-Pael-R, 1:200, monoclonal anti-TH, 1:1000 (Chemicon), anti-NeuN, 1:200 (Chemicon) or anti-CNPase, 1:100 (Sigma). Secondary antibodies were conjugated to either Alexa 488 or Alexa 546 (Molecular Probes) and viewed with a laser scanning confocal microscope system (Fluoview, Olympus).
Polyclonal Anti-Pael, or anti-BiP (N-20) (both from Santa Cruz)were used for immunocytochemistry on transfected SH-Sy5Y cells.
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New Substrate for Parkin Links Disease to ER Stress
