Alternatives to Laemli?

Wolfgang Schechinger hubahopp at gmx.de
Mon Feb 16 18:05:44 EST 2004


I remember that a german company named Carl Roth used to sell a phosphate
based laemmli claiming phosphate had a smaller temperature coefficient and
thus yielded a more stable pH during the boiling step and thus prevented
some chemical proteolysis (to reduce the number of 'artificial' bands).

If someone is interested, I'll dig out the receipe.

Wo


At 16:57 16.02.2004 -0600, EK wrote:
>
>> have relatively equal mobilities through low percentage PAAG. However,
>when
>> both reach separating gel at pH 8.8, glycine picks up creating a charge
>> "vacuum" zone immediately behind the transition line between the two gel
>
>---- I will correct myself: its "immediately below the transition line" (in
>vertical systems)
>
>> layers. This charge vacuum slows down migration of proteins as they enter
>> the separating gel, causing a stacking effect. I hope I remember that
>> correctly.
>>
>> As for the original post, I don't think SDS-based buffer would work for
>> studying protein/RNA complexes at all, simply because SDS would denature
>the
>> protein. There are non-denaturing buffer systems, like Tris-borate-EDTA
>> (TBE) that work fine for that purpose.
>>
>>
>
>
>
Wolfgang Schechinger


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