Good day, I prepare Laemmli buffer 2X from Bio-Rad by adding 950uL of Laemmli buffer + 50uL of B-mercaptoethanol for a final volume of 1mL for SDS-PAGE. Since a lot of Laemmli buffer still remains, can it be re-used for future samples? or at least for running gel electrophoresis in the same week?
If so, at what temperature is it stored and for how long?
You can keep laemmli sample buffer in freeze condition. We always store the sample buffer in -20 degrees in small aliquots (in 2 mL microtube)
Hope it help!
Dear Eduardo
the Leamnil with out reducing agent is stable for long time but once you add b-mercapto or DTT, it has short stability at 4°C or RT. As Mabrur suggest, you can make aliquotes and store it at -20°C for long time.
If you have some Laemil with B-mercapto that was stored at RT or 4°C you can add again B-mercapto end use it. But you cannot do it many times because any addiction of the reducing will dilute the Laemil and at the end you risk that it is not work properly.
good luck
Manuele
I agree with previous posts. Just one additional hint: DTT (Cleland's reagent, doi:10.1021/bi00892a002) has a redox potential that is better for reducing Cys-Cys bonds than BME. Stinks less, too.
We do make 1x Laemmli buffer and store at 4 degree. Just before the use, we aliquot and add beta mercaptoethanol. The amount that remains can be used within a week not more than that and that too stored at 4 degree.
Hope it helps.
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