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Position: Discussion forums > Bioscience biotechniques discussion forums > Immunology forum and cytometry forum
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ELISA with fresh frozen plasma
Posted by: biogirl (IP Hidden, New member, 2)
Date: May 6, 2005 04:17PM
hello all,
i'm trying to run an ELISA using fresh frozen plasma (FFP). but upon thawing out FFP, cryoprecipitate forms which totally throws my readings off. has anyone else had this problem? any ideas on troubleshooting? i've tried thawing at 4C vs. 37C, filtering, centrifuging, everything. but nothing seems to really work. any help would be much appreciated. thanks!!
Re: ELISA with fresh frozen plasma
Posted by: mygateway (IP Hidden, Junior member, 10)
Date: May 9, 2005 10:08AM
First of all, the shelf life for FFP is around 1 year. Secondly, thawing FFP slowly in at 4c will result in cryoprecipitate which formed by some plasma protein factors.
Re: ELISA with fresh frozen plasma
Posted by: femmeauburn (IP Hidden, Advanced member, 115)
Date: May 12, 2005 10:11AM
Slowly thaw the FFP at 1-6C. A cryoprecipitate will form that can be removed by subsquent centrifugation to pellet the cryoprecipitate. Filtering should also remove the cryoprep from the thawed FFP. You must let the precipitate form before trying to remove it.
Re: ELISA with fresh frozen plasma
Posted by: liangma (IP Hidden, New member, 1)
Date: August 3, 2005 06:21PM
I'm planning to use ELISA/Western to detect bacterial-specific antibodies in frozen plasma from infected patients. I'm looking for information regarding possible problems/disadvantages of using plasma (rather than serum) for serologic work. I'd be very grateful if anyone could give me any suggestions/advice on this.
As mentioned above, cryoprecipitation and shorter shelf life are the major problems? Is it possible to convert the frozen plasma to serum? Thank you. Liangma
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