Following treatment with GGA or other compounds, HuH-7 cells grown on glass inserts in a 24-well plate were rinsed with Ca-free PBS (PBS(-); Sigma Aldrich) and fixed for 40 min with 4% paraformaldehyde containing 2% sucrose in PBS(-), and then rinsed with PBS(-). Cells were then permeabilized with 0.5% Triton X-100 and nonspecific binding blocked with 10% FBS. Next, cells were incubated at 4°C overnight with polyclonal anti-XBP1s antibody, followed by 2.5 h incubation with Alexa-488-labeled goat anti-rabbit IgG antibody (Invitrogen, Molecular Probes, Tokyo, Japan). After rinsing with PBS(-), cells were mounted in PermaFluor (Beckman Coulter, Brea, CA, USA), covered on a glass slide, and observed under a confocal laser-scanning fluorescence microscope, LSM700 2Ch URGB equipped with Axio Observer Z1 Bio (Carl Zeiss, Göttingen, Germany).
Lsm700 2ch urgb
Manufactured by Zeiss
Sourced in Germany
The LSM700 2Ch URGB is a laser scanning microscope system developed by Zeiss. It is designed for high-resolution imaging and analysis of biological samples. The system features two detection channels and can capture images in the ultraviolet, red, green, and blue color spectrum.
Automatically generated - may contain errors
Lab products found in correlation
Axio observer z1 bio, by Zeiss (2 mentions)
Permafluor, by Beckman Coulter (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Alexa 488 labeled goat anti mouse igg antibody, by Thermo Fisher Scientific (1 mentions)
Anti p53 monoclonal antibody, by Cell Signaling Technology (1 mentions)
2 protocols using lsm700 2ch urgb
1
Immunofluorescence Analysis of XBP1s in HuH-7 Cells
2
Immunofluorescence Analysis of p53 in Liver Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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After 20 μM GGA or ethanol treatment, HuH-7 and PLC/PRF/5 cells on a glass insert in a 24-well plate were rinsed with PBS(−) and fixed for 40 min with 4% paraformaldehyde containing 2% sucrose in PBS(−) and then rinsed with PBS(−). The cells were permeated with 0.5% TritonX-100 and blocked with 10% FBS. The cells were then incubated at 4°C overnight with a monoclonal anti-p53 antibody (Cell Signaling Technology), followed by a 2.5-h incubation with an Alexa-488-labeled goat anti-mouse IgG antibody (Invitrogen, Molecular Probes, Tokyo, Japan). After rinsing with PBS(−), the cells were mounted in ParmaFluor (Beckman Coulter, Brea, CA, USA), covered on a slide glass, and observed under a confocal laser-scanning fluorescence microscope, an LSM700 2Ch URGB equipped with Axio Observer Z1 Bio (Carl Zeiss, Göttingen, Germany).
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