ELISA with bacterially expressed protein
marthoe at biologie.uni-freiburg.de
Fri Nov 10 07:23:52 EST 2000
I want do an ELISA with a (Drosophila) protein expressed in bacteria.
The protocol I've found in Current Protocols in Mol Biol (p 11.2.18,
Preparation of bacrerial cell lysate antigens)
is in short:
- resuspend bacterial pellet (from 1ml liquid culture) in 1ml 10mM HEPES
- add 0.2ml lysozyme sol., incubate 5min on ice
- centrifuge and resuspend pellet in 1.2ml TEN (Tris/EDTA/NaCl, pH 7.2)
- add 0.065ml 10% SDS, incubate 10min at 37degrees.
according to the protocol the samples are READY FOR ELISA at this point.
But unfortunately nothing is said about how to dilute the samples. Or
should I use them undiluted?
any help would be appreciated,
Institut fuer Biologie III
Labor Prof. Fischbach
Email marthoe at biologie.uni-freiburg.de
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