800 C3H/10T1/2 cells were seeded per well in a 96 well plate (glass bottom culture plates, MatTek, PBK96G-1.5-5-F) and kept at 37-degree Celcius for 24 h. Cells were fixed with 4% formaldehyde in PBS for 10 min at room temperature. Cells were permeabilized with 0.2% digitonin (EMD Millipore, 300410) in PBS (Corning, 21-040-CV) for 10 min at room temperature. Next, the cells were incubated in 3% BSA (in PBS, Sigma, #9418) for blocking for 1 h at room temperature followed by incubation in primary antibody (ATRX, Santa Cruz, sc-15408, 1:200) at 4°C overnight, followed by secondary antibody (1:600, Invitrogen, A32754) incubation at room temperature for 1 h. Cells were washed three times with PBS for 5 min followed by DAPI staining (2 μg/ml in PBS, Sigma, D9564) for 5 min at room temperature. Mounting media (Vectashield, H-1000) was added immediately after DAPI staining. Cells were imaged using Widefield Microscope CellDiscoverer7 (CD7) automated widefield high-throughput system (Zeiss). Images were processed with ImageJ software (http://rsb.info.nih.gov/ij/ ). For the ATRX antibody, the ImageJ Brightness/Contrast was set as 30/112. For DAPI, the Brightness/Contrast was set as 37/123. All images were processed with the same parameter settings for each antibody.
Widefield microscope celldiscoverer7 cd7
Manufactured by Zeiss
The Widefield Microscope CellDiscoverer7 (CD7) is a high-performance microscope system designed for cell culture and live-cell imaging applications. It features a widefield illumination system and advanced optics for capturing high-quality images of living cells.
Automatically generated - may contain errors
2 protocols using widefield microscope celldiscoverer7 cd7
1
Immunofluorescence Staining of ATRX in 3D Cell Cultures
2
Quantifying ATRX Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
800 C3H/10T1/2 cells were seeded in a 96 well plate (glass bottom culture plates, MatTek, PBK96G-1.5-5-F) and keep cells in 37-degree for 24 h. Cells were fixed with 4% formaldehyde in PBS for 10 minutes at room temperature. Cells were permeabilized with 0.2% digitonin (EMD Millipore, 300410) in PBS (Corning, 21-040-CV) for 10 minutes at room temperature. Next, the cells were incubated in 3% BSA (in PBS, Sigma, #9418) for blocking for 1 h at room temperature followed by incubation in primary antibody (ATRX, Santa Cruz, sc-15408, 1:200) at 4-degree overnight, followed by secondary antibody (1:600, Invitrogen, A32754) incubation at room temperature for 1 h. Cells were washed three times with PBS for 5 minutes followed by DAPI staining (2 μg/ml in PBS, Sigma, D9564) for 5 minutes at room temperature. Mounting media (Vectashield, H-1000) was added immediately after DAPI staining. Cells were imaged using Widefield Microscope CellDiscoverer7 (CD7) automated widefield high-throughput system (Zeiss). Images were processed with ImageJ software (http://rsb.info.nih.gov/ij/ ). For the ATRX antibody, the ImageJ Brightness/Contrast was set as 30/112. For DAPI, the Brightness/Contrast was set as 37/123. All images were processed with the same parameter settings for each antibody.
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!