Blank control (Black line): Raji (Black).
Primary Antibody (green line): Rabbit Anti-Leptinreceptor antibody (SL0109R)
Dilution: 1μg /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% ice-cold methanol overnight at 4℃ and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. 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: K562.
Primary Antibody (green line): Rabbit Anti-Leptin receptor antibody (SL0109R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1μg /test.
Protocol
The cells were 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.
Cell: (mo)Hepatocyte(2% BSA at 4°blocked for 30 minutes.1% Paraformaldehyde fixed for 30 minutes,0.1%TritonX-100 permeablized for 2 minutes).
Concentration:1:100.
Incubation: 40 minutes.
Host/Blank: Hepatocytes.
Flow cytometric analysis of Rabbit Anti-Leptin receptor(long)/AF488 Conjugated antibody (SL0109R-AF488)(green) compared with control in the absence of primary antibody (blue) followed by (mo)hepatocytes.
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