Blank control (blue line): Hela (blue). Primary Antibody (green line): Rabbit Anti-phospho-PKC delta (Tyr52) antibody (orb6749), dilution: 1 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): F (ab)2 fragment goat anti-rabbit IgG-FITC, dilution: 1 µg/Test. Protocol, The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.
Paraformaldehyde-fixed, paraffin embedded (rat lymphoid), 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 (phospho-PKC delta (Tyr52)) Polyclonal Antibody, Unconjugated (orb6749) at 1:200 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat ovary), 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 (phospho-PKC delta (Tyr52)) Polyclonal Antibody, Unconjugated (orb6749) at 1:200 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
Sample: Cerebellum (Rat) Lysate at 40 ug, Primary: Anti-phospho-PKC delta (Tyr52) (orb6749) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 77 kD, Observed band size: 80 kD.
Tissue/Cell: mouse brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37C for 20 min, Incubation: Anti-phospho-PKC delta (Tyr52) Polyclonal Antibody, Unconjugated (orb6749) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining.
Immunohistochemical staining of Mouse brain tissue using PKC delta (phospho-Tyr52) antibody.
Immunohistochemical staining of Mouse brain tissue using PKC delta (phospho-Tyr52) antibody.
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