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Excess betamercaptoethanol during sample preparation


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4 replies to this topic

#1 criscastells

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Posted 14 February 2013 - 12:50 AM

We had a problem during Western Blot sample preparation. The charge buffer was prepared with an excess of betamercaptoethanol (1:1) instead of (5:100). May it be a problem? And why?

Thanks

#2 mdfenko

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Posted 14 February 2013 - 04:56 AM

should not be a problem (other than greater than usual odor).

you could correct it by increasing the volume with the other components until all are of the correct concentration (then aliquot and freeze so that it doesn't all go bad at the same time).

Edited by mdfenko, 14 February 2013 - 04:58 AM.

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genius does what it must
i do what i get paid to do

#3 criscastells

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Posted 15 February 2013 - 02:56 AM

Thanks!

I can not correct the concentrations since I finished the western. I wanted to know it in order to take into account the results or not.

#4 mdfenko

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Posted 15 February 2013 - 04:53 AM

there should be no significant difference (unless your normal sample buffer has an inadequate amount of bme) but you should confirm the result by repeating the blot (maybe with both normal and excess bme).

Edited by mdfenko, 15 February 2013 - 04:55 AM.

talent does what it can
genius does what it must
i do what i get paid to do

#5 criscastells

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Posted 15 February 2013 - 05:20 AM

Thanks you very much for your help!




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