Cross banding on Westerns using monoclonal Ab? help!
Mr. A.F. Hill
ahill at crc.ac.uk
Tue Jun 1 11:19:45 EST 1993
i am trying to perform some western blots on some proteins extracted from
E.coli which contains an expression vector for a protein we wish to study
(ie to use as an assay for expression). i am using a monoclonal antibody for
the primary and either a horseradish-peroxidase conjugated secondary antibody
(use ECL to develop) or a alkaline phosphatase conjugated secondary antibody
(NBT & BCIP to develop).
the problem is that several bands develop on the membrane - including bands
from the molecular weight standards that were loaded onto the SDS-PAGE gel.
is there any reason why several bands appear - obviously unrelated to the
protein of interest, and which shouldn't (i presume) react with the monoclonal
any help would be appreciated...
Andy Hill |
Prion Disease Group | E-Mail: ahill at uk.ac.crc
Dept. Biochemistry & Molecular Genetics |-----------------------------------
St Mary's Hospital Medical School | Phone: 071 723 1252 ext. 5469
LONDON W2 1PG | Fax: 071 706 3272
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