[IF=2.173] Liu B et al. Effects of Autophagy on Synaptic-Plasticity-Related Protein Expression in the Hippocampus CA1 of a Rat Model of Vascular Dementia. Neurosci Lett. 2019 Jun 1;707:134312.  WB ;  Rat.  

PubMed:31163225

Tissue/cell: human glioma tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min; Incubation: Anti-LC3 α/MAP1A/MAP LC3 Alpha/Beta Polyclonal Antibody, Unconjugated (bs-1534R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Blank control (blue line): Hela (blue). Primary Antibody (green line): Rabbit Anti-MAP1LC3A antibody (bs-1534R) Dilution: 0.2μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE Dilution: 1μg /test. Protocol The cells were fixed with 70% ethanol (Overmight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min at -20℃.Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X 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 20,000 events was performed.

Blank control: RAW264.7. Primary Antibody (green line): Rabbit Anti-MAP1LC3A antibody (bs-1534R) Dilution: 2ug/Test; Secondary Antibody : Goat anti-rabbit IgG-FITC Dilution: 0.5ug/Test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature.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 20,000 events was performed.

Blank control: RAW264.7. Primary Antibody (green line): Rabbit Anti-MAP1LC3A antibody (bs-1534R) Dilution: 2ug/Test; Secondary Antibody : Goat anti-rabbit IgG-FITC Dilution: 0.5ug/Test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature.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 20,000 events was performed.