For immunofluorescence assays, 200,000 cells were plated on SecureSlip silicone supported coverglasses (Sigma Aldrich) in 6-well plates that had been pre-coated for 60 minutes with 0.01 mg/mL human fibronectin (Calbiochem) in PBS. The following day, cells were fixed in 4% paraformaldehyde (PFA) diluted in PBS for 15 minutes at room temperature. Cells were permeabilized with 0.2% Triton X-100 in PBS for 10 minutes. Blocking was performed in 2.5% normal goat serum blocking solution (Vector Laboratories). Cells were incubated in primary MAGOH/MAGOHB antibody (Abcam, ab38768, rabbit, 1:200) for 1 hr at RT. A Cy-3 conjugated anti-rabbit secondary (Abcam, ab97075, 1:200) and DAPI (1:1000, Life Technologies) were then used for 1 hr at RT. Cells were mounted and imaged on an Axio Observer fluorescent microscope (Zeiss) using AxioVision software (Zeiss). Nuclear/Cytoplasmic ratio was quantified by Image J. Nuclear outlines were determined based on DAPI signal. Cytoplasmic signal was defined as signal in the whole cell minus signal within the nuclear area.
Secureslip silicone supported coverglasses
Manufactured by Merck Group
SecureSlip silicone supported coverglasses are a type of laboratory equipment designed to cover and protect sample slides. They feature a silicone coating that securely adheres to the slide surface, preventing the coverglass from slipping or shifting during use.
Automatically generated - may contain errors
Lab products found in correlation
Axio observer fluorescent microscope, by Zeiss (4 mentions)
Ab38768, by Abcam (2 mentions)
Ab97075, by Thermo Fisher Scientific (2 mentions)
Normal goat serum blocking solution, by Vector Laboratories (2 mentions)
Axiovision software, by Zeiss (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
2 protocols using secureslip silicone supported coverglasses
1
Immunofluorescence Assay for MAGOH/MAGOHB
2
Immunofluorescence Assay for MAGOH/MAGOHB
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunofluorescence assays, 200,000 cells were plated on SecureSlip silicone supported coverglasses (Sigma Aldrich) in 6-well plates that had been pre-coated for 60 minutes with 0.01 mg/mL human fibronectin (Calbiochem) in PBS. The following day, cells were fixed in 4% paraformaldehyde (PFA) diluted in PBS for 15 minutes at room temperature. Cells were permeabilized with 0.2% Triton X-100 in PBS for 10 minutes. Blocking was performed in 2.5% normal goat serum blocking solution (Vector Laboratories). Cells were incubated in primary MAGOH/MAGOHB antibody (Abcam, ab38768, rabbit, 1:200) for 1 hr at RT. A Cy-3 conjugated anti-rabbit secondary (Abcam, ab97075, 1:200) and DAPI (1:1000, Life Technologies) were then used for 1 hr at RT. Cells were mounted and imaged on an Axio Observer fluorescent microscope (Zeiss) using AxioVision software (Zeiss). Nuclear/Cytoplasmic ratio was quantified by Image J. Nuclear outlines were determined based on DAPI signal. Cytoplasmic signal was defined as signal in the whole cell minus signal within the nuclear area.
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!