Overlay Peak curve showing PC-3 cells stained with CSB-RA056302A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/1*106cells) for 45min at 4°C. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4°C.Control antibody (green line) was Rabbit IgG (1ug/1*106cells) used under the same conditions. Acquisition of >10,000 events was performed.
Immunofluorescence staining of Hela cell with CSB-RA056302A0HU 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 4C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L) .
IHC image of CSB-RA056302A0HU diluted at 1:100 and staining in paraffin-embedded human glioma 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 4C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB.
IHC image of CSB-RA056302A0HU 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 4C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB.
Western Blot Positive WB detected in: SH-SY5Y whole cell lysate(30µg) , HEK293 whole cell lysate(30µg) , Hela whole cell lysate(30µg) , MCF-7 whole cell lysate(30µg) , K562 whole cell lysate(30µg) , Jurkat whole cell lysate(30µg) , NIH/3T3 whole cell lysate(30µg) , Rat brain tissue lysate(30µg) All lanes: Citrate synthase antibody at 1:1000 Secondary Goat polyclonal to rabbit IgG at 1/40000 dilution Predicted band size: 52 kDa Observed band size: 52 kDa Exposure time:20s
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