Hi,
I would like to avoid some inespecific bands in my western. I have cell lysis western a it is full of bands. Could somebody tell me how I can avoid the inespecific bands? I tried with less incubation, and with other antibodies but I couldnīt do it.
thanks a lot
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2 replies to this topic
#2
mdfenko
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Posted 09 September 2010 - 07:50 AM
you may need to change your blocking agent and/or antibody concentration.
have you determined whether the problem is your primary or secondary antibody? i suspect the problem is with your secondary antibody because you have the problem with all of your primary antibodies. try diluting it more (and make sure you have blocking agent in the solution).
have you determined whether the problem is your primary or secondary antibody? i suspect the problem is with your secondary antibody because you have the problem with all of your primary antibodies. try diluting it more (and make sure you have blocking agent in the solution).
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
#3
SUVEAR
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Posted 10 September 2010 - 01:07 AM
thank you,
I know that the problem is with my secondary antibody. I changed it but I have the same problem I will tried with diferent concentration and bloking agent. I will tried with BSA.
Thank you
I know that the problem is with my secondary antibody. I changed it but I have the same problem I will tried with diferent concentration and bloking agent. I will tried with BSA.
Thank you
Edited by SUVEAR, 10 September 2010 - 01:08 AM.
