Immunofluorescence staining of PC-3 cell with CSB-PA621886ESR1HU at 1:25, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and and permeated by 0.2% TritonX-100 for 15 min. Then 10% normal goat serum to block non-specific protein-protein interactions . 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).
Immunohistochemistry image of CSB-PA621886ESR1HU diluted at 1:50 and staining in paraffin-embedded human prostate 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: HL-60 whole cell lysate(20µg), Hela whole cell lysate(20µg), Jurkat whole cell lysate(20µg), PC-3 whole cell lysate(20µg) All lanes: CHRNA2 antibody at 1:1000 Secondary Goat polyclonal to rabbit IgG at 1/40000 dilution Predicted band size: 59 kDa Observed band size: 60 kDa Exposure time: 300s
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