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Hi, I was looking through the protocol for your Plasminogen activator Inhibitor Type 1 Human Chromogenic Activity Kit (ab108894) and while you add exogenous tPA to serum & plasma samples, this is NOT suggested for cell culture supernatants. Is this a mistake?
Dr. Mark Fowler, Senior Cell Biologist, Unilever R&D Colworth
B50, Colworth Science Park, Sharnbrook, Bedford, UK, MK44 1LQ
Tel: +44 (0) 1234 222993
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Asked on Jan 18 2018
Thank you for getting in touch.
I can confirm that the tPA standard must be added to all the sample including cell culture supernatant for the Plasminogen activator Inhibitor Type 1 Human Chromogenic Activity Kit (ab108894).
I am very sorry about this mistake in our protocol booklet. Thank you that you brought it to our attention and our team is going is updating it at the moment.
Below is the information from the lab:
"The tPA standard need to be added for all the sample since the PAI activity will be evaluated by inhibit the tPA activity. You could add your cell culture sample without dilution to see how much they can inhibit the tPA activity. Ideally, you will do a series dilution of your cell culture sample to see how the dilution will affect the tPA activity and so that you can see the full picture of the inhibition."
Since the amount of tPA standard to be added to cell culture supernatant is not strictly determined, you might need to do some pilot experiments to find the best condition.
Please do not hesitate to let me know if you have any other questions.
Answered on Jan 18 2018