Anti AKT1, phospho Ser473, Recombinant Antibody  

Western Blot
Positive WB detected in 293 whole cell lysate(treated with Calyculin A or not)
All lanes Phospho-AKT1 antibody at 1.08μg/ml
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 60 KDa
Observed band size: 60 KDa

IHC image of CSB-RA001553A473phHU diluted at 1:100 and staining in paraffin-embedded human breast cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

IHC image of CSB-RA001553A473phHU diluted at 1:100 and staining in paraffin-embedded human lung cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

Immunofluorescence staining of Hela cells(treated with 100mM EGF for 20min) with CSB-RA001553A473phHU at 1:68,counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4℃. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG (H+L).

Immunoprecipitating Phospho-AKT1 in 293 whole cell lysate treated with Calyculin A
Lane 1: Rabbit control IgG(1μg)instead of CSB-RA001553A473phHU in 293 whole cell lysate treated with Calyculin A.
For western blotting,a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: CSB-RA001553A473phHU(3μg)+ 293 whole cell lysate treated with Calyculin A(1mg)
Lane 3: 293 whole cell lysate treated with Calyculin A (20μg)