Blank control (black line): HepG2 (black) (The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with PBST for 30 min on room temperature), Primary Antibody (green line): Rabbit Anti-LDLreceptor antibody (orb10972), Dilution: 1 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 µg/Test.
Blank control: A431. Primary Antibody (green line): Rabbit Anti-LDL receptor antibody (orb10972), 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, A431 cells were fixed with 4% PFA (10 min 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 20000 events was performed.
Blank control: Raw264.7. Primary Antibody (green line): Rabbit Anti-LDL receptor antibody (orb10972), 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 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 20000 events was performed.
HepG2 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37C for 20 min, Antibody incubation with (LDL receptor) polyclonal Antibody, Unconjugated (orb10972) 1:100, 90 minutes at 37C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
Sample: Human colon Lysate at 45ug, Primary: Anti-LDL receptor (orb10972) at 1:300 dilution, Secondary: HRP conjugated Goat Anti-Rabbit IgG (orb572747) at 1:5000 dilution, Predicted band size: 92kD, Observed band size: 92kD.
Sample: K562 (Human) Cell Lysate at 30 ug, Raw264.7 (Mouse) Cell Lysate at 30 ug, HepG2 (Human) Cell Lysate at 30 ug, SW480 (Human) Cell Lysate at 30 ug, Primary: Anti-LDL receptor (orb10972) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 92 kD, Observed band size: 120 kD.
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