Western blotting analysis using NUMB antibody (Cat#65751). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with NUMB antibody (Cat#65751, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:50,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using NUMB antibody (Cat#65751). NUMB expression in wild type (WT) and NUMB shRNA knockdown (KD) HT-1080 cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with NUMB antibody (Cat#65751, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Immunocytochemical staining of HT-1080 cells with NUMB antibody(Cat#65751, 1:1,000). Nuclei were stained blue with DAPI; NUMB was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol. Scale bar, 20 μm.
Flow cytometric analysis of NUMB expression in HT-1080 cells using NUMB antibody (Cat#65751, 1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol, chill on -20℃ before use. Green, isotype control; red, NUMB.
Validation of NUMB knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HT-1080 cells were stained with NUMB antibody (Cat#65751, 1:2,000) and analyzed using BD flow cytometer. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol, chill on -20℃ before use.
