Troubleshooting Guide: Western Blot
|
Troubleshooting Guide: Western Blot
From R&D systems. It gives troubleshooting guide on several common problems on western blot: low protein transfer, high background, low protein binding, multiple bands staining etc.
Last update 31-May-2007, Rating Poor of 8 votes.
|
Write your comment
|
Dear Richa,
I am a bigginer in western Blot Myself,but as far as I know,A+B solutions have short lifetime after using on the membrane ,so ,after an hour u have to use them on your membrane again.
best,
Mitra.
Rating: n/a
Reply
|
|
|
I am doing western blot of protein. And my problem is that when i develop using ECL reagent from my lab the blot does not work. Butif i borrow the reagent from neighbouring lab it works. So I changed the reagent kit, film and tried again the same probloem persists. Could the dveloping maching have something to do with this. some developing temperature difference or something? Can some one hel me with this pleasre
Rating: n/a
Reply
|
|
|
Hey Syed Wasim, maybe your sample is too small of an amount. You can try concentrating your sample on a Speed Vac first, then loading it onto your gel.
Rating: n/a
Reply
|
|
|
i am doing western blotting for quite some time but am getting faint signal only, although i made some changes (transfer time, current, changing Pab conc )but was not able to get desired result when i develope it with DAB, please help
Rating: n/a
Reply
|
|
|
I am in the loading process of the western blot and why is the loading buffer getting all cloudy when it was running. what could I have done differently? thank you
Rating: n/a
Reply
|
|
|
i have problem about power.power has error 4
Rating: Excellent!
Reply
|
|
|
I am in the loading process of the western blot and why is the loading buffer getting all cloudy when it was running. what could I have done differently? And would this cloudiness affect the result when i am ready to expose? Thank you very much.
Rating: n/a
Reply
|
|
|
i am doing western blotting with HRP secondary and ECL. the proplem i am facing is unusual. when i develo blot for 10min some bands (specific) get highlighted in some lanes on the X-ray film and in another half an hour exposure bands (specific) in other lanes highlighted but the previous band are not there?? why so
please help
Rating: n/a
Reply
|
|
Related resource
Western blot stripping protocol
Semi-dry western blot transfer procedure
Homemade Chemiluminescent Protocols?
Western-Blot: PBS/TBS
Western blot problem
Western blot protocol
large protein transfer efficiency for western blot
Western blot
|
