Hi
I used the expression vector pCDNA3.1/flag to subcloning the C-terminal region of BRCA2 and transfect it into BRCA2 deficient cells. After Immunoprecipitation and Western blotting analysis with a flag antibodies, I didn't see the expected band of approximately 80 KD but two bands of 50 and 30 KD.
Any suggestions?
Thanks in advance
Laura
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4 replies to this topic
#2
GNANA
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Posted 22 December 2011 - 03:20 AM
did you run the Antibody along with the IP, just to make sure your 50KD is not the Ab chain.....
I would prefer being perfectionist rather than a passionist in Research.
I always had an alternate hypothesis....
I always had an alternate hypothesis....
#3
Lauretta77
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Posted 22 December 2011 - 05:20 AM
No, I didn't! Thank you for your suggestion, I'll verify it.
Just an information, how many antibodies I have to run?
Just an information, how many antibodies I have to run?
#4
GNANA
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Posted 23 December 2011 - 08:31 AM
i usually denature the same amount i use for IP.....may be u can also use bit less than that...
I would prefer being perfectionist rather than a passionist in Research.
I always had an alternate hypothesis....
I always had an alternate hypothesis....
#5
Lauretta77
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Posted 26 December 2011 - 03:29 AM
Ok!
Thank you very much!
Thank you very much!
