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Silver stained 2-d gel BLOB!!!

Silver stained 2-d gel BLOB!!! - 2-D Gel Electrophoresis Forum

Silver stained 2-d gel BLOB!!! - 2-D Gel Electrophoresis Forum. A forum dedicated to two dimensional gel electrophoresis, discussion about systems, and troubleshooting its problems.


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Old 04-22-2010, 04:44 PM
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Default Silver stained 2-d gel BLOB!!!



Hi
I was wondering if someone could help me. Everytime I run my protein samples on 2-D gels and silver stain these gels I get this large blob at the base of the gel (see attached picture). I use GE IEF strips and ampholytes and Invitrogen pre-cast gels. I also use Invitrogen LDS sample buffer. I take great care not to touch the gels before staining.
Any help appreciated.
sam
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Old 04-22-2010, 05:09 PM
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Default Re: Silver stained 2-d gel BLOB!!!

Looks like the gels are broken from rough handling.
I bought a new rubber spatula and brought it to the lab, to help me handle these big gels.

Might try that.

Plus rinse gel well before staining. Make sure stain isn't old.
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Old 07-06-2010, 06:17 AM
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Default Re: Silver stained 2-d gel BLOB!!!

Hi

To identify a problem initialy u can stain with comassie blue and followed by silverstaining, so that u could able to identify is that due to handling or sample difussion.
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Old 03-18-2011, 12:58 PM
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Default Re: Silver stained 2-d gel BLOB!!!

Hi sam_bookoo,
i also had the same problem in our last gel stained with silver stain........what was your sample, mine was cyanobacterial protein
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Old 05-30-2011, 06:44 AM
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Default Re: Silver stained 2-d gel BLOB!!!

Hi,
Please use fresh and pure reagents (SDS running buffer, all reagents used for 2DE, staining solutions, as danfive mentioned). Poor quality reagents can produce this kind of blob.
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Old 06-02-2011, 04:19 PM
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Default Re: Silver stained 2-d gel BLOB!!!

Hi Quys, I found that this blob was due to an excess in carrier ampholytes!!!
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