HepG2.2.15 cells were grown on coverslips and treated with R837 as stated earlier. The cells were washed thrice with Phosphate buffered saline (PBS) and then fixed with 4% Paraformaldehyde (Sigma) in PBS for 10 min at room temperature (RT). Cells were washed thrice with PBS and permeabilized with 1% Triton X-100 in PBS for 10 min at RT, washed with PBS thrice followed by blocking with 3% BSA in PBS for 1 h at RT. Cells were incubated with Anti-NFКB (eBioscience) for 1 h at RT, washed thrice with PBST (PBS + 0.05% Tween20) and then incubated with secondary antibody (Alexa-Fluor anti-rabbit 488, Invitrogen) for 1 h in dark at RT, followed by three washes with PBST. Coverslips were mounted with mounting media containing DAPI (Sigma Aldrich). Fluorescence for Alexa and DAPI was visualized with Nikon Ti-E confocal microscope with A1RMP scanner head equipped with Nikon imaging software (NIS).
A1rmp scanner head
Manufactured by Nikon
The A1RMP scanner head is a component of Nikon's imaging systems. It serves as a key part of the scanning mechanism, responsible for accurately and precisely capturing high-quality images or data. The A1RMP scanner head operates using established scanning principles, but the specific technical details and intended applications are not included in this factual description.
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Lab products found in correlation
3 protocols using a1rmp scanner head
1
Analyzing NF-κB Activation in HepG2.2.15 Cells
2
Immunofluorescence Visualization Protocol
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Immunofluorescence was carried out as described previously45 (link). Cells were fixed with 4% paraformaldehyde or methanol, permeabilized with 1% Triton X-100 or 100% methanol, blocked with 3% BSA, and stained with the indicated antibodies. Coverslips were mounted after staining with DAPI (4′,6-diamidino-2-phenylindole) and photographed using a Nikon T1E confocal microscope with an A1RMP Scanner Head.
3
Quantifying NF-κB Activation in LPS-Treated HepG2.2.15 Cells
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HepG2.2.15 cells were grown on coverslips and treated with 4 μg/mL of LPS-B5 Ultrapure as stated earlier. The cells were washed with phosphate buffered saline (PBS) three times and then fixed with 4% Paraformaldehyde (Sigma) in PBS for 10 min at room temperature (RT). Cells were washed thrice with PBS and permeabilized with 1% Triton X-100 in PBS for 10 min at RT, washed with PBS thrice followed by blocking with 3% BSA in PBS for 1 h at RT. Cells were incubated with Anti-NF-κB [1:1000 (e-Bioscience14-6731-81) for 1 h at RT, washed thrice with PBST (PBS + 0.05% Tween20) and then incubated with secondary antibody (Alexa-Fluor anti-rabbit 488 (1:1000), Invitrogen) for 1 h in dark at RT, followed by three washes with PBST. Coverslips were then mounted with mounting media containing DAPI (Sigma Aldrich). Fluorescence for Alexa and DAPI was visualized with Nikon Ti-E confocal microscope with A1RMP scanner head equipped with Nikon imaging software (NIS).
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