Technique / Proteomics and protein biochemistry / Western blot assay
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 of 9 votes.
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By kapil shah
on 07-Nov-2008
I am planning to seperate insect proteins on a fluorotrans pvdf membrane, but I notice that the proteins do not transfer to the membrane and remain in the acrlamide gel itself. I am using 20% methanol in the transfer buffer. Kindly point out where am I going wrong. I electroblot at 30V for 16-18 h. |
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By Mitra
on 09-Jul-2008
Dear Richa, |
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By Vinita
on 08-May-2008
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 |
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By splorch
on 02-May-2008
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. |
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By syed wasim
on 08-Apr-2008
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 |
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By rghosh
on 17-Jan-2008
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 |
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By sobhan
on 29-Nov-2007
i have problem about power.power has error 4 |
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By maria
on 04-Sep-2007
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. |
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By richa
on 31-May-2007
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 |
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