Custom Monoclonal Antibody Production

Western Blot
Positive WB detected in:Hela whole cell lysate,HEK293 whole cell lysate
All lanes : PODXL antibody at 2.5 ug/ml
Secondary
Goat polyclonal to Mouse IgG at 1/5000 dilution
Predicted band size: 59KDa, 56KDa
Observed band size: 59KDa

Western Blot
Positive WB detected in:U251 whole cell lysate,SH-Sy5y whole cell lysate
All lanes :NES antibody at 3 ug/ml
Predicted band size: 260KDa
Observed band size: 260KDa

IHC image of A antibody 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 SP system.

Overlay histogram showing THP-1 cells stained with CD31 (red line) at 1:500. The cells were incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4℃.The secondary antibody used was FITC goat anti-mouse IgG (H+L) at 1/200 dilution for 1 h at 4℃. Isotype control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.

Immunoprecipitating PODXL in HEK293 whole cell lysate
Lane 1: Rabbit monoclonal IgG(1 ug)instead of PODXL in HEK293 whole cell lysate. For western blotting,a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: PODXL(8 ug)+ HEK293 whole cell lysate(500 ug)
Lane 3: HEK293 whole cell lysate (10 ug)

Immunofluorescence staining of A549 cells with A antibody at 1:100, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C.The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG (H+L).