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

Western blot
All lanes: ELAVL1 antibody at 6µg/ml + A431 whole cell lysate
Secondary
Goat polyclonal to rabbit IgG at 1/10000 dilution
Predicted band size: 37, 39 kDa
Observed band size: 37 kDa

Western Blot
Positive WB detected in: HEK293 whole cell lysate(30μg), U-251MG whole cell lysate(30μg), PC-3 whole cell lysate(30μg), HL-60 whole cell lysate(30μg), HepG2 whole cell lysate(30μg), COLO205 whole cell lysate(30μg), HeLa whole cell lysate(30μg), Jurkat whole cell lysate(30μg)
All lanes: ELAVL1 antibody at 1:1000
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Exposure time: 10s

IHC image of CSB-RA555528A0HU diluted at 1:100 and staining in paraffin-embedded human prostate 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 Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB.

Immunofluorescence staining of Hela cell with CSB-RA555528A0HU at 1:50, 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).

Overlay Peak curve showing MCF-7 cells stained with CSB-RA555528A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/1*10⁶cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/1*10⁶cells) used under the same conditions. Acquisition of >10,000 events was performed.