IHC image of PHO8727 diluted at 1:100 and staining in paraffin-embedded human cervical 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°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

IHC image of PHO8727 diluted at 1:100 and staining in paraffin-embedded human kidney tissue 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°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated ABC system.

Immunofluorescence staining of Hela cells with PHO8727 at 1:100, 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°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).

Immunoprecipitating FLOT1 in Hela whole cell lysate
Lane 1: Rabbit control IgG instead of PHO8727 in Hela whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: PHO8727 (8μg) + Hela whole cell lysate (500μg)
Lane 3: Hela whole cell lysate (10μg)

Immunoprecipitating FLOT1 in K562 whole cell lysate
Lane 1: Rabbit control IgG instead of PHO8727 in K562 whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/5000)
Lane 2: PHO8727 (8μg) + K562 whole cell lysate (500μg)
Lane 3: K562 whole cell lysate (20μg)

Western Blot
Positive WB detected in: HepG2 whole cell lysate, K562 whole cell lysate, Hela whole cell lysate, Rat brain tissue, Mouse brain tissue
All lanes: FLOT1 antibody at 3μg/ml
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 48, 43 kDa
Observed band size: 48 kDa