Subcellular Protein Fractionation Kit for Cultured Cells

Protein isolation Mammalian cells - HOG

Experiment
Protein isolation Mammalian cells - HOG
Product
Subcellular Protein Fractionation Kit for Cultured Cells from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
- Instructions are followed regarding the manufacturer protocol, except fractions corresponding to cytosol, membrane and nuclei were boiled 5 min at 95°C in the presence of Laemmlli sample buffer

Publication protocol

Cells were seeded onto 10 cm2 plastic Petri dishes and cell fractionation was done according to manufacturer. Briefly, attached cells were washed three time with ice-cold PBS++ and then scraped in ice-cold PBS. Cells were transferred in microcentrifuge tubes and pelleted by centrifugation at 500 × g for 5 min. Supernatants were eliminated and cell pellets were resuspended in ice-cold cytosolic extraction buffer (CEB, Pierce). After 10 min at 4°C, tubes were centrifuged at 500 × g for 5 min. Supernatants (corresponding to cytosolic fractions) were collected. Pellets were resuspended in membrane extraction buffer (MEB, Pierce), vortexed and left 10 min at 4°C. Tubes were then centrifuged at 3000 × g for 5 min. Supernatants (corresponding to membrane fractions) were collected and pellets were resuspended in nuclear extraction buffer (NEB, Pierce), giving the nuclear fraction. Fractions corresponding to cytosol, membrane and nuclei were boiled 5 min at 95°C in the presence of Laemmlli sample buffer. Fractions were then subjected to SDS-PAGE on 10% polyacrylamide gels and transferred onto nitrocellulose membranes.

Full paper   Login or join for free to view the full paper.

Reviews

Subcellular Protein Fractionation Kit for Cultured Cells from Thermo Fisher Scientific has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein isolation Mammalian cells - HOG using Subcellular Protein Fractionation Kit for Cultured Cells from Thermo Fisher Scientific.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Subcellular Protein Fractionation Kit for Cultured Cells below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Protein isolation Mammalian cells - HOG using Subcellular Protein Fractionation Kit for Cultured Cells from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms