Anti BACE  

Immunocytochemistry of BACE antibody. Cells: 3T3 cells. Fixation: formaldehyde and blocked with 10% serum for 1 h at RT. Antigen retrieval: heat mediation with a citrate buffer (pH6). Primary antibody: BACE antibody at 10 μg/mL overnight at 2-8°C. Secondary antibody: Peroxidase goat anti-rabbit secondary antibody at 1:250 for 45 min at RT. Localization: BACE is localized in the cell membrane, endoplasmic reticulum, Golgi apparatus, endosome, and the cell surface. Counter stained with Hematoxylin.

Immunofluorescence of Rabbit Anti-BACE Antibody. Cells: NIH/3T3 Cells. Fixation: 4% Paraformaldehyde. Primary Antibody: Anti-BACE at 20μg/mL. Secondary Antibody: Goat anti-Rabbit at 1:500. Staining: DAPI. Cytosol and membrane staining.

Immunofluorescence of Anti-BACE Antibody. Immunolabeling patterns of BACE1 expression and the lenti-siRNA distribution. Sections from APP transgenic mice treated with the eGFP-tagged lenti siRNA vectors (green) were co-immunolabeled with an antibody against BACE1 (red) and imaged with the LSCM. All sections are from the hippocampus of treated mice. (a？c) Lenti-siBACE1-6？treated mice. Areas within the hippocampus expressing the eGFP-tagged vector have reduced BACE1 immunolabeling. (d？f) Mice treated with the eGFP-tagged control lenti-siGlut4 show unchanged expression of BACE1 in the hippocampus. (g？i) Mice treated with a saline vehicle show unchanged expression of BACE1 in the hippocampus. (Singer et al., 2005).

Immunofluorescence of Anti-BACE Antibody. Decreased BACE1 expression in DRG following siRNA3 transfection. DRG neurons were transfected with 1 μg siRNA3 plasmid and incubated for 48 hours in 37°C. DRG neurons were stained for BACE1 using the Anti-BACE antibody. (a,b) Neurons transfected with the control plasmid pSUPER-EGFP (green) did not display any changes in BACE1 expression (red). (c,d) DRG neurons transfected with siRNA3 displayed reduced BACE1 expression in the axon. (Hyun, 2007).

Immunohistochemistry of Rabbit anti-BACE Anitbody. Tissue: Mouse Brain. Fixation: Formaldehyde blocked with 10% serum for 1hr at RT. Antigen retrieval: heat mediation with citrate buffer pH6. Primary Antibody: Anti-BACE at 2.5μg/mL overnight at 2-8°C. Secondary Antibody: goat anti-Rabbit IgG HRP at 1:250. Counterstain: hematoxylin.

Immunohistochemistry of Rabbit Anti-BACE Antibody. Characterization of the effects of lenti-siBACE1-6 expression in the brains of APP transgenic mice. (a？d) Anti-eGFP immunoreactivity in the hippocampus (the injection site) shows comparable and consistent expression of lenti-siRNA constructs in the dentate gyrus (dg) and stratus polymorphus (sp). (e) Anti-BACE1 immunoreactivity in the hippocampus of nontransgenic mice treated with lenti-siGlut4. (f) Reduced BACE1 immunostaining in the hippocampus of nontransgenic mice treated with lenti-siBACE1-6 vector. (g) Intense BACE1 immunoreactivity in the hippocampus of APP transgenic mice treated with lenti-siGlut4. (h) Reduced BACE1 expression in APP transgenic mice treated with lenti-siBACE1-6 vector. (i,j) Anti-BACE1 reacted with pyramidal cell bodies in the neocortex, which was not injected. (Singer et al., 2005).

Western Blot of BACE antibody. Lane 1: Human brain tissue lysate. Lane 2: Human brain tissue lysate with the presence of blocking peptide. Lane3: Mouse 3T3 cell lysate. Load: 35 μg per lane. Primary antibody: BACE antibody at 1 μg/mL for overnight at 4°C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: 55.7 kDa, 76 kDa for BACE. Other band(s): BACE splice variants and isoforms.

Western Blot of Anti-BACE Antibody. Loading: 15 μg of lysates per lane. Primary Antibodies: Anti-BACE 200-401-Z18 (1μg/mL), BACE (1μg/mL), beta-actin (1μg/mL), and GAPDH (0.02μg/mL), for 1hr at RT in 5% NFDM/TBST. Secondary Antibody: Goat anti-rabbit IgG HRP conjugate at 1:10,000.

Western Blot of Anti-BACE Antibody. Wildtype and BACE −/− MEFs were exposed to HNE (15_M) for 2 hr. BACE1 levels were examined by Western blot with anti-BACE antibodies. (Jo et al., 2010)

Western Blot of Anti-BACE Antibody. Lanes 1？4 are frontal cortex homogenates from human and mouse brains. Lane 1 is from a neurologically unimpaired aged human control case, lane 2 from a BACE1-deficient mouse, lane 3 from a nontransgenic mouse and lane 4 from hBACE1 transgenic mouse. Lanes 5？7 are lysates from HEK293T cells transfected with a plasmid vector expressing eGFP, mBACE1 and hBACE1, respectively. BACE1 antibody recognized both the mouse and human forms of BACE1. (Singer et al., 2005).