Tuesday:We continued the pull down assay with the proteins and bead that I put on to incubate overnight. We loaded the flow through samples on to one gel and the beads on to the other two gels.
The stained flow through
gel appeared to have a lot less protein than before…. which is good it means that
more proteins have bound to the beads this time.The other 2 gels with the elution and bead samples were transferred to a nitrocellulose membrane for western blotting.
While the gel was running we prepared 2 more SDS-Page gels. Once transferred, the membrane was put in blocking overnight.
We then decided to optimise
the beads used for the pull down and used a mixture of amylose beads for one
protein and nickel beads on an identical protein in order to compare them. We
ran all of the samples on a gel.
Today was very
challenging-phew!!
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