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western blot sample in high salt concentration


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6 replies to this topic

#1 szang

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Posted 23 January 2010 - 10:50 AM

Hi ereryone

My samples are in different salt conentration(0.15M Nacl, 0.5M Nacl, 1.0M Nacl). When i do western to identify them, but i found some bands disappear and some ugly bands. It is Especially strange that the band also disappears in the normal salt concentration.

Is it because the salt concentration affects the result? Should i dilute the sample with water?

Does anyone have apinions on it, please tell me. Thank you very much

#2 mdfenko

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Posted 25 January 2010 - 07:32 AM

salt will have an effect on the running of your samples.

rather than dilute you can dialyze the samples (by drop dialysis) to reduce the salt concentration.
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#3 szang

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Posted 25 January 2010 - 01:24 PM

salt will have an effect on the running of your samples.

rather than dilute you can dialyze the samples (by drop dialysis) to reduce the salt concentration.


Thanks, i repeat the experiment and it works.

#4 DNAze

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Posted 14 July 2011 - 09:56 AM

salt will have an effect on the running of your samples.

rather than dilute you can dialyze the samples (by drop dialysis) to reduce the salt concentration.




Can you do drop dialysis on samples already in laemmli buffer?

#5 mdfenko

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Posted 14 July 2011 - 10:20 AM

Can you do drop dialysis on samples already in laemmli buffer?


yes. dialyze against laemmli buffer.
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#6 DNAze

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Posted 14 July 2011 - 01:11 PM


Can you do drop dialysis on samples already in laemmli buffer?


yes. dialyze against laemmli buffer.



I'm doing my own search currently, but do you by chance happen to have a link or a protocol? that would be greatly helpful.

#7 mdfenko

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Posted 15 July 2011 - 06:08 AM

the attached protocol is for nucleic acids but is similar to drop dialysis for proteins. also, you can use standard dialysis membrane instead of filter disks.

in the olden days, we used to dialyze our samples for sds-page against sample buffer (without bromphenol blue or glycerol, which were added after dialysis) prior to loading.

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talent does what it can
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