To visualize the nuclear translocation of NF-κB in ALL-SIL cells, isogenic control and venetoclax-resistant cells were plated on poly-L-lysine coated coverslips placed in a 6-well plate. After 12 to 16 hours, the cells were fixed for 15 minutes with 4% paraformaldehyde at room temperature and washed three times with PBS. Then they were blocked in 5% goat serum with 1 mg/mL BSA and 0.3% TritonX-100 for 2 hours. Rabbit anti-human NF-κB p65 antibody (D14E12, Cell Signaling Technology) was used to stain cells overnight at 4°C. The following day, cells were washed three times with PBS for 5 minutes, and A488-conjugated goat anti–rabbit secondary antibody (Invitrogen) was applied for 1 hour at room temperature. After secondary antibody incubation, cells were washed three times with PBS for 5 minutes each and counterstained with DAPI for 5 minutes. Afterward, cells were washed three times with PBS for 5 minutes each. The coverslips were mounted using anti-fade fluorescence mounting medium (Abcam, ab104135), and cells were imaged using a Leica TCS SP8 Confocal/Multi-Photon high-speed upright microscope. Nuclear to cytoplasmic NF-κB p65 ratio was calculated using ImageJ.
Tcs sp8 confocal multi photon high speed upright microscope
Manufactured by Leica
The Leica TCS SP8 is a high-speed, upright confocal and multi-photon microscope. It is designed for advanced imaging applications that require high-resolution and fast image acquisition.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using tcs sp8 confocal multi photon high speed upright microscope
1
Visualizing NF-κB Translocation in ALL-SIL Cells
2
Visualizing CD55 and Lipid Rafts in CSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To visualize the expression and localization of CD55 and cholera toxin B, a lipid raft marker, A2780 and TOV112D CSCs were plated on coverslips placed in a 6-well plate. After 12–16 h, the cells were fixed for 15 min with 4% paraformaldehyde at room temperature and washed three times with PBS. After washing, cells were incubated with A488-conjugated cholera toxin B (Invitrogen) for 15 min and washed again three times. Then they were blocked in 5% goat serum with 1 mg/ml BSA for 2 h. Mouse monoclonal CD55 antibody (Santa Cruz) was used to stain cells overnight at 4°C. The following day, cells were washed three times with PBS for 5 min, and A647-conjugated goat anti–mouse secondary antibody (Invitrogen) was applied for 1 h at room temperature. After secondary antibody incubation, cells were washed three times with PBS for 5 min each and counterstained with DAPI for 5 min. Afterward, cells were washed three times with PBS for 5 min each. The coverslips were mounted using 50% glycerol, and cells were imaged using a Leica TCS SP8 Confocal/Multi-Photon high-speed upright microscope.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!