As my cells are adherent breast cancer cell lines I don't want to trypsinize the cells and want to study the expression of some hormonal receptors as well as some cytokines by using confocal microscopy.
I read few protocols but they used to trypsinize and do flowcytometry based detection of cytokines and the same an be used for confocal detection.
But I am looking for a protocol in which i can do intracellular staining without any dissociation procedure.
Hey Jaykumar, here is a protocol taht may help you;
Good Luck :)
I Jaykumar,
in my following paper you will find all staining (cell, tissue, immunofluorescence and immunohistocehmical staining). I hope it will help
- PLoS One. 2010 Feb 2;5(2):e9023. doi: 10.1371/journal.pone.0009023.
The high affinity IgE receptor Fc epsilonRI is expressed by human intestinal epithelial cells.
Untersmayr E1, Bises G, Starkl P, Bevins CL, Scheiner O, Boltz-Nitulescu G, Wrba F, Jensen-Jarolim E.
- http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0009023
If you any question please contact me
Good luck
Giovanna
I definitively recommend PLA (proximity Ligations Assay). With PLA you can detect a single protein or interaction between 2 proteins. Just grow your cells on slide (check the link) .Here below you can see 2 links . you might find what you need.
Good luck
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2868571/
http://www.sigmaaldrich.com/catalog/product/sigma/z694576?lang=en®ion=SE
http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma-Aldrich/General_Information/1/duolink-short-protocol.pdf
- Badges
- Science topic