Cells were grown on glass coverslips and were fixed 72 h posttransfection using ice-cold 8:2 methanol:acetone for 15 min at 4 °C and blocked with 2% BSA-PBS. Coverslips were incubated with anti-human PANX1 antibody (1:500; PANX1 CT-412; 0.35 μg/μl) and anti-mouse β-catenin (1:450 BD Biosciences# 610153). Nuclei was stained with Hoechst 33342 (1:1000), Alexa Fluor 647 anti-mouse and 555 goat anti-rabbit IgG (2 mg/ml, 1:500) were used as secondaries, and mounted using Airvol (Mowiol 4-88; Sigma Aldrich) prior to imaging. Immunofluorescence images were obtained using a Zeiss LSM 800 Confocal Microscope with a Plan-Apochromat 63x/1.40 Oil DIC objective (Carl Zeiss).
Airvol mowiol 4 88
Manufactured by Merck Group
Airvol (Mowiol 4-88) is a polyvinyl alcohol (PVOH) product manufactured by Merck Group. It is a white to off-white, free-flowing powder.
Automatically generated - may contain errors
2 protocols using airvol mowiol 4 88
1
Immunofluorescence Staining of PANX1 and β-Catenin
2
Immunofluorescence Analysis of PANX1 Expression
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Cells were grown on glass coverslips and were fixed 48–72 h posttransfection using ice-cold 8:2 methanol:acetone for 15 min at 4°C and blocked with 2% BSA-PBS. Coverslips were incubated with anti-human PANX1 antibody (1:500; PANX1 CT-412; 0.35 μg/μl), Hoechst 33342 (1:1000), and Alexa Fluor 488 goat anti-rabbit IgG (2 mg/ml, 1:700) and mounted using Airvol (Mowiol 4-88; Sigma Aldrich) prior to imaging. Immunofluorescence images were obtained using a Zeiss LSM 800 confocal microscope with a Plan-Apochromat 63×/1.40 Oil DIC objective (Carl Zeiss, Oberkochen, Germany). The laser lines used include 405 nm (Hoechst 33342) and 488 nm (Alexa488).
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