19-23FEB2018

I broke some more IRM12 cells and cehcked the A280nm for checking the sonication. The A280nm of the lysate was 35  which means that the sonication have worked well. I do an AKTA purification but now using a buffer A with no Imidazole and a buffer B with 500mM as before and also using a new His resin column 1ml. I run the AKTA fractions and dialysed the fractions where I saw the UV pick (see gel on page         ). The protein was detected but with a lot of unspecific binding.

I put the fractions (2 and 3) for dialysis but then the weather got snowy and left the dialysis for 5 days in the cold room

05MAR

I measured the A280nm of the before dialysis (AKTA fractions) protein: 3.523
                                                  after dialysis (AKTA fractions) protein: 8.443
                                                  dialysate (liquid in which I was making the dialysis, buffer A):0.0305

I spun down the dialysed protein (3900rpm, 10min) and saw a pellet forming. I run on the gel the pellet (discard the supernatant and add water), the supernatant, and the dialysate.
See gel on page