1 reply to this topic

[Joon Kim]

I'm preparing membranes from CHO cells cultured in Corning 75cm² flasks. My protocol for this is generic: Scrape cells at 90~100 confluence, homogenise and spin at 60,000g. My final pellet is resuspended in 3ml of buffer.

But when I got to my Bradford assay to determine protein concentration, I'm finding that from one flask, I get around 0.5mg/ml. Although I resuspend my pellet in 3ml, which might be high, I'm not entirely convinced that this is all I get out of it?

I was wondering if anyone has experience in this, and may have an idea of how much protein one may expect to get?

Thank you for your time.

[Curtis]

This doesn't look like a good protocol to me. usually fractionation protocols have more centrifugation steps. we have many protocols at the main protocol-online.org page. That's where I found my own protocol.