[IF=4.718] Arumugam P et al. Expression of a Functional IL-2 Receptor in Vascular Smooth Muscle Cells. The Journal of Immunology,2018 ji1701151.  WB ;  Human.  

PubMed:doi:10.4049/jimmunol.1701151

Sample: Raji(Human) Cell Lysate at 30 ug Primary: Anti-IL2 Receptor beta (bs-1959R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 58 kD Observed band size: 58 kD

Paraformaldehyde-fixed, paraffin embedded (human lung carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (IL2 Receptor beta) Polyclonal Antibody, Unconjugated (bs-1959R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control: U937 (blue). Primary Antibody:Rabbit Anti-IL2 Receptor beta antibody(bs-1959R), 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-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-1959R, 1μg /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.