Thursday:
I feel like I have
written hundreds of these blog entries now and at this point you would think I
would be bored doing the same things every day but when things don’t work,
which it turns out yesterday's western blot was not successful, then we get to
try again making small changes as we go.
I completed all 6 pull
down assays from Monday again but this time with a lot more amylose beads,
hopefully something will stick this time meaning that the western blot might
actually work.
All of the samples were
prepared for the four gels which we will use tomorrow, I look forward to seeing
if it will work. (Cross your fingers for me.)
While preparing each
sample with the help of one of my colleagues, she somehow managed to spill half
a millilitre of SDS loading dye onto the bench top (if only you could have seen
her face, first shock and then a naughty smile), at which point she decided to
try and clean it up by pipetting it with a 10ul tip (instead of wiping it up),
the two of us trying very hard not to laugh, but the tip made little dent in
the little purple ‘puddle’.
The samples that the
volunteers prepared from yesterday’s pull down assay were run on 3 gels and
transferred to a membrane, normal WB protocol was carried out. All of the membranes
were stripped of their antibodies using stripping buffer (clever name!!) and
left in blocking buffer overnight. We are going to probe these tomorrow with
different antibodies.
Today was a little
confusing but we got there in the end.
No comments:
Post a Comment