Sample:
A549 Cell (Human) Lysate at 30 ug
Primary: Anti-GPR78(bs-1807R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 39 kD
Observed band size: 39 kD
Blank control: 293T Cells(blue).
Primary Antibody: Rabbit Anti-GPR78/AF488 Conjugated antibody (bs-1807R-AF488), Dilution: 1 µg in 100 µL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG/AF488(orange) ,used under the same conditions.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) . The cells were washed twice with 1 X PBS. The cells were incubated in 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions followed by the incubated with antibody (bs-1807R-AF488, 1 µg /1x10^6 cells) for 30 min on ice. Acquisition of 20,000 events was performed.
Dilution: Blank control (blue line): U937 (blue).
Primary Antibody (green line): Rabbit Anti-GPR7 antibody (bs-1807R)
Dilution: 1 µg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 µg /test.
Protocol
The cells were fixed with 2% paraformaldehyde for 10 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.
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