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Blank control: SH-SY5Y. Primary Antibody (green line): Rabbit Anti-MAP2 antibody (orb11455), Dilution: 1 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF647, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed. |
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Paraformaldehyde-fixed, paraffin embedded (Human brain glioma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (MAP2) Polyclonal Antibody, Unconjugated (orb11455) at 1:400 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining. |
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Paraformaldehyde-fixed, paraffin embedded (Mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (MAP2) Polyclonal Antibody, Unconjugated (orb11455) at 1:400 overnight at 4C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868805) for 90 minutes, and DAPI for nuclei staining. |
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Paraformaldehyde-fixed, paraffin embedded (Mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (MAP2) Polyclonal Antibody, Unconjugated (orb11455) at 1:400 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining. |
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Paraformaldehyde-fixed, paraffin embedded (Rat brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (MAP2) Polyclonal Antibody, Unconjugated (orb11455) at 1:400 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining. |
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Sample: Cerebrum (Mouse) Lysate at 40 ug, Primary: Anti-MAP2 (orb11455) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 201 kD, Observed band size: 280 kD. |
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Tissue/Cell: rat brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Blocking buffer (normal goat serum) at 37C for 20 min, Incubation: Anti-MAP2/MAP-2a.b.c Polyclonal Antibody, Unconjugated (orb11455) 1:200, overnight at 4C, The secondary antibody was Goat Anti-Rabbit IgG, FITC conjugated (orb868805) used at 1:200 dilution for 40 minutes at 37C. DAPI (5 ug/ml, blue) was used to stain the cell nuclei. |