8 Questions 22 Answers 0 Followers
Questions related from Madison Edwards
I would like to do a dot blot assay to test for a negatively charged protein (pI
01 October 2019 2,317 3 View
I am trying to purify a protein with a large intrinsically disordered domain, and I have been able to increase the solubility of the protein by adding an MBP-tag on the N-terminus. The protein is...
17 February 2019 8,501 20 View
I would like to purify my MBP-tagged protein by ammonium sulfate precipitation followed by an amylose bead column. I could perform a dialysis step in the middle, but if the binding to amylose...
13 January 2019 5,827 5 View
I would like to purify a protein using the IMACT system where my protein has an intein and chitin-binding domain. I cannot use salt in my purification. The protocol says that you can use 50-1000...
29 August 2018 7,804 3 View
I have a MBP fusion protein that is not binding to amylose beads. I have already tried adding glucose to the media (although I did not add it to my starter culture) and I am not using detergents...
23 August 2018 5,871 7 View
I have a protein that binds RNA and has lots of disorders regions. I have tried purifying from E. coli and baculovirus/sf9 systems. In both, the protein appears to aggregate and crash out of...
20 July 2018 5,119 10 View
I am purifying a 74 kDa protein (about half of the protein is predicted to be disordered) that has an His6-Maltose-binding protein tag (~118 kDa total) total. After eluting from the nickel beads I...
26 April 2018 9,274 6 View
I am purifying a 118 kDa fusion protein (His6-maltose binding protein-my protein). I see induction of a protein at 118 kDa, but also multiple other products are induced that are 75-100 kDa in...
21 December 2017 1,576 8 View
