I treated cells with stearic acid for different times. Collect total protein lysate after 3-time wash with PBS. Test protein concentration with Bio-Rad DC Protein Assay Reagents Package Kit. Although same ug protein is loaded into each well, GAPDH or Actin blot signal is not consistent. But control treatment with BSA has no problem. What is the problem? Thanks for your help.
Thank you so much, Anwar! If that is true, maybe the salt form of stearic acid is better. I will go to try.
I don't think there will be a chemical reaction between stearic acid and amino groups. My guess is that the stearic acid acted as a detergent, causing loss of soluble cytoplasmic proteins from the cells due to membrane solubilization.
Thank you, Adam. I am trying to use higher cell density and check the data.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Proteins