Sample: Muscle (Mouse) Lysate at 40 ug
Primary: Anti- Adiponectin Receptor 2 (SL0611R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 44 kD
Observed band size: 44 kD
Sample:
Lane 1: Heart (Mouse) Lysate at 40 ug
Lane 2: Skeletal muscle (Mouse) Lysate at 40 ug
Lane 3: Heart (Rat) Lysate at 40 ug
Lane 4: Skeletal muscle (Rat) Lysate at 40 ug
Lane 5: 293T (Human) Cell Lysate at 30 ug
Primary: Anti-Adiponectin receptor 2 (SL0611R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 44 kD
Observed band size: 43 kD
Sample:
293T(Human) Cell Lysate at 30 ug
Primary: Anti-Adiponectin receptor 2 (SL0611R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 44 kD
Observed band size: 48 kD
Sample:
muscle (Mouse) Lysate at 40 ug
Primary: Anti-Adiponectin receptor 2 (SL0611R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 44 kD
Observed band size: 44 kD
Sample:
Lung (Mouse) Lysate at 40 ug
Primary: Anti-Adiponectin receptor 2 (SL0611R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 44 kD
Observed band size: 46 kD
Tissue/cell: human rectal 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-Adiponectin Receptor 2 Polyclonal Antibody, Unconjugated(SL0611R) 1:300, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Tissue/cell: rat pancreas 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-Adiponectin Receptor 2 Polyclonal Antibody, Unconjugated(SL0611R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Blank control: HepG2(blue). Primary Antibody:Rabbit Anti-Adiponectin receptor 2 antibody (SL0611R,Green); Dilution: 1μg 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-FITC(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde for 10 min at 37℃. Primary antibody (SL0611R, 0.2μg /1x10^6 cells) were incubated for 30 min at room temperature, 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/FITC antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 40 min at room temperature. Acquisition of 20,000 events was performed.
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