Sample:
Jurkat(Human) Cell Lysate at 30 ug
Primary: Anti-CD32  (bs-2573R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 31 kD
Observed band size: 31 kD

Sample: K562 Cell Lysate at 30 ug
Primary: Anti- CD32 (bs-2573R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 31 kD
Observed band size: 33 kD

Blank control: U937 (blue).
Primary Antibody:Rabbit Anti-CD32 antibody(bs-2573R), Dilution: 0.2μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions );
Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) . Primary antibody (bs-2573R, 0.2μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.