ELISA with bacterially expressed protein

Martin Hoehne marthoe at biologie.uni-freiburg.de
Fri Nov 10 07:23:52 EST 2000


Hello everybody,

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,

Martin

 
______________________________________
Martin Hoehne
Institut fuer Biologie III
Labor Prof. Fischbach
Schaenzlestr. 1
79104 Freiburg

Tel +49-(0)761-203-2759
Fax +49-(0)761-203-2866 
Email marthoe at biologie.uni-freiburg.de






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