| |||||||||
| Login :: Register :: Search forums :: Top Users :: Reagent |
|
Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
|
western blot problem
Posted by: Gugo (IP Hidden, New member, 5)
Date: January 25, 2008 02:47AM
Hi all of you,
I did a western blot and I obtained a weak signal, so I blocked the membrane again and re-incubated primary and secondary antibodies using a less dilution. The signal dissapeared. I did an stripping and reprobing using a more sensitive ECL system, but no signal again. Any of you could know what happened? My samples are valuable and it is a pity I have to throw it away. I would very apreciate your help. Thank you!
Re: western blot problem
Posted by: HCscientist (IP Hidden, New member, 3)
Date: January 26, 2008 12:58AM
There are several possible reasons: 1) You did not completely remove stripping buffer, which inactivate your antibody. Wash enough before re-blocking. 2) During ECL development, I wash too much and get rid of signal. Do it as soon as you can. PerkinElmer sell WesternLighting reagent, cheaper price and better quality. Try it.
Re: western blot problem
Posted by: Gugo (IP Hidden, New member, 5)
Date: February 5, 2008 03:11AM
Hi HCscientist,
Thank you for your advice, but it didn't work. I don't know what happened. I will think about it... Thank you again
|
We are moving ... Please post to our new community forums
|
|
||||||||