I am studying gene expression profile through analyses of proteins expressed upon treatment of cancer stem cell line with a novel herbal substance (in the extract) to predict gene function in the mechanism of reaction. I would, therefore, need to know the best method to determine/screen proteins expressed solely consequent upon this treatment in achieving this considering;
1. Model of the study (cancer stem cell line)
2. Testing molecules (drug candidate from herb)
3. Analyses involved.
Hi Kamoru,
You could try SILAC/AACT (stable isotope labeling with amino acids in cell culture/amino acid-coded tagging ) followed by LC-MS/MS (liquid chromatography- tandem mass spectrometer). This approach is not really expensive and will give a great vision of what would happen to cells that treated vs control. For more information, check below link.
Article Systematic Proteomic Identification of the Heat Shock Protei...
Hi Kamoru
this will be rather expensive but I guess you want to go for RPPA (e.g. Article The Use of Reverse Phase Protein Arrays (RPPA) to Explore Pr...
), most important hits should be verified by Western blot,complemented by multi color immune fluorescence imaging to get an idea of homogeneity in the culture.
Best
Heiko
Heiko Dussmann I am very grateful for this informative direction. Thank you, Sir.
Hi Kamoru,
You could try SILAC/AACT (stable isotope labeling with amino acids in cell culture/amino acid-coded tagging ) followed by LC-MS/MS (liquid chromatography- tandem mass spectrometer). This approach is not really expensive and will give a great vision of what would happen to cells that treated vs control. For more information, check below link.
Article Systematic Proteomic Identification of the Heat Shock Protei...
Hello
The SILAC/ACT followed by LC-MS/MS it's a good suggestion.
Good Luck
The answers are educative, thank you Profs Saleh Alkarim , Ahmed Dhamad and Heiko Dussmann
- Badges
- Science topic
- Similar topics
- Chemistry
- Pharmacology
- Pharmaceuticals
- Drugs