(Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.

Flow cytometric analysis of calpain 2 expression in HT-1080 cells using calpain 2 antibody. Green, isotype control; red, calpain 2.

Immunocytochemical staining of HT-1080 cells with Calpain 2 antibody. Nuclei were stained blue with DAPI; Calpain 2 was stained magenta with Alexa Fluor? 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Scale bar, 20 μm.

Western blotting analysis using calpain 2 antibody. Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with calpain 2 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively.

Western Blot
Positive WB detected in: 293T whole cell lysate
All lanes: CAPN2 antibody at 3.4µg/ml
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 80, 72 kDa
Observed band size: 80 kDa

IHC image of CSB-PA004496LA01HU diluted at 1:300 and staining in paraffin-embedded human tonsil 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.

IHC image of CSB-PA004496LA01HU diluted at 1:300 and staining in paraffin-embedded human appendix 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.

Immunofluorescence staining of HepG2 cells with CSB-PA004496LA01HU 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 CAPN2 in Hela whole cell lysate
Lane 1: Rabbit control IgG instead of CSB-PA004496LA01HU in Hela whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: CSB-PA004496LA01HU (8µg) + Hela whole cell lysate (500µg)
Lane 3: Hela whole cell lysate (10µg)