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About antibodies


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2 replies to this topic

#1 sorcener

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Posted 17 November 2009 - 09:52 AM

Hi everybody,

I'm starting a new lab and been having problems with a western blot. I have tryed everything and I have no signal!

People here told me once the lab fridge went out of current a whole weekend and all antibodies are there! My boss says nothing happened, that abs can resit that, but now I'm not so sure (luminol was there too by the way).

I did dot blot just with my secondary on PVDF membrain, (1:10 and 1:10 000) and saw nothing =(

What do you think about it?

#2 bob1

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Posted 17 November 2009 - 03:40 PM

It depends on the Ab, some are more stable than others. Ideally antibodies should be stored aliquotted and frozen and only thawed once or twice. However, most antibodies will be OK at 4 deg C for a month or so, after that they usually start to degrade. They are not very stable at room temperature, but would probably last a couple of days.

The luminol should be fine, though the H2O2 is not so stable.

#3 aimikins

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Posted 20 November 2009 - 03:51 PM

I would not expect the blotting to work until you can do the control with your secondary and get a signal. it does seem that your detection reagents are more likely to expire than your antibodies and perhaps you can start there?
"it is a miracle that curiosity survives formal education" -A.E.




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