Protein: Huh7(human) lysates at 30ug;
Primary: Anti-LXR alpha (SL10311R) at 1:500;
Secondary: HRP conjugated Goat-Anti-Rabbit IgG(bse-0295G-HRP) at 1: 5000;
ECL excitated the fluorescence;
Predicted band size : 46 kD
Observed band size : 46 kD
Protein: liver(mouse) lysates at 40ug;
Primary: Anti-LXR alpha (SL10311R) at 1:300;
Secondary: HRP conjugated Goat-Anti-Rabbit IgG(bse-0295G-HRP) at 1: 5000;
ECL excitated the fluorescence;
Predicted band size : 46 kD
Observed band size : 46 kD
Tissue/cell: Rat testis 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,SLC0005) at 37℃ for 20 min;
Incubation: Anti-LXR alpha Polyclonal Antibody, Unconjugated(SL10311R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-LXR alpha antibody (SL10311R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at 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:THP-1.
Primary Antibody (green line): Rabbit Anti-LXR alpha antibody (SL10311R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. 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.
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