Hi.
I'm working on MMPs. When I did dot blots I recived great signal but when I tried when I tried western blots there was no signal at all. I don't know whats wrong cause I'm using the same conditions as in dot blots. I checked my transfer and it's ok.
What else might be wrong?
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#2
mdfenko
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Posted 23 April 2010 - 12:09 PM
dot blots are usually of the native protein, western blots are of denatured protein. your antibody may not work with the denatured protein. if it is a commercial antibody, does the data sheet say that the antibody is certified for western blot?
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genius does what it must
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#3
Alsai
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Posted 27 April 2010 - 04:17 AM
Yes, antibodies are for western blot.
mdfenko, on Apr 23 2010, 09:09 PM, said:
dot blots are usually of the native protein, western blots are of denatured protein. your antibody may not work with the denatured protein. if it is a commercial antibody, does the data sheet say that the antibody is certified for western blot?
#4
mdfenko
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Posted 27 April 2010 - 07:16 AM
the amount of protein in the band may be significantly lower than in the dot blot. you may need to incubate longer with and/or use a higher concentration of the antibody.
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
