hello everyone!
HeLa cells were seeded in 6-well plates at 200,000 cells per well. They were left for 2–3 days to reach 70–80% confluency. Then, determination of protein concentration was performed with BCA. but I think the protein concentration I measured is too low. What should be the expected protein concentration for HeLa cells (seeded in a six-well plate)?
any recommendations are appreciated!
thanks in advance!
Hello Naile Merve Güven
The expected protein concentration will vary depending on many factors. In such a case in order to achieve maximum protein yield, you should try to optimize your protocol.
I have provided some suggestions to a similar question on ResearchGate platform. You could incorporate those suggestions in your protocol too to obtain maximum yield of protein.
https://www.researchgate.net/post/Protein_extraction_from_SH-SY5Y_neuroblastoma_cells_and_differentiated_neurons
Best.
I know that the amount of protein can vary from cell to cell. The protocol I follow is similar to yours. I want to know approximately what the amount I expect to achieve at the end of all this should be. so that I know whether I am following the right protocol or that a change needs to be made.
Thank you again, have a good day!
Dear Nalie
Regarding the previous reports, approximately 150-200 pg total protein per (HeLa) cell is expected. So, given the initial cell seeding and the average doubling time, you can calculate the expected value in your case.
Hi Naile
Estimated protein yield for HeLa cell line is 200 µg / 10^6 cells.
You may want to refer to the link attached below.
https://research.fredhutch.org/paulovich/en/typical-protein-yields.html
Best.
The protein concentration of HeLa cells, or any cell type, can vary based on several factors such as cell density, growth conditions, and the specific experimental context. Typically, protein concentration is measured in micrograms or milligrams of protein per milliliter of cell lysate.
If you are working with HeLa cells and need to determine the protein concentration, you can follow these general steps:
It's important to note that the specific protocol and reagents used for protein quantification may vary based on the assay chosen. Additionally, for accurate quantification, it's advisable to perform the assay in triplicate or according to the recommended procedure for the chosen method.
Keep in mind that the protein concentration can also vary based on the purpose of your experiment. For example, if you are preparing samples for Western blotting, you may use a different lysis buffer and quantification method compared to preparing samples for mass spectrometry.
Always refer to the specific protocols provided by the manufacturers of the reagents and kits you are using, as they may have detailed instructions tailored to their products.
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