I have a plan to investigate cell signaling on NKG2D-expressing CD4. For this matter, I plan to isolate primary cells from patients and use western blot technique to measure the expression level of phosphorylated and total proteins related to my particular signaling. However, within our body, the number of NKG2D-expressing CD4 cells is very low. Due to the small number of cells, it seems not possible for me to do western blot. Is there any alternative way to measure protein expression from small number of cells?
Yes, you can measure protein expression as well as phosphorylation status of protein by Flowcytometry. It will be easier and cheaper method than WB. For FCM you need not to sort cells.
Feel free for further help regarding experiment designing to interpretation.
Sharing like of a not for advantage of flowcytometry over WB.
https://blog.cellsignal.com/when-should-i-use-flow-cytometry-for-signaling-instead-of-western-blot
Yes, you can measure protein expression as well as phosphorylation status of protein by Flowcytometry. It will be easier and cheaper method than WB. For FCM you need not to sort cells.
Feel free for further help regarding experiment designing to interpretation.
Sharing like of a not for advantage of flowcytometry over WB.
https://blog.cellsignal.com/when-should-i-use-flow-cytometry-for-signaling-instead-of-western-blot
yes, you could perform flow cytometry to measure phosphorylation of intracelullar proteins in rare cell populations
Thank you much Jitendra Kumar Shandilya and Luis Ibarra . Is there any suggestion the way how to set up experiment by using flowcytometry for cell signaling?
Tell me the signals you want to check. What signalling molecule you want to see?
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