21 Questions 20 Answers 0 Followers
Questions related from David Vizarraga
Hello, my name is David I want to study the inhibitory effect of the pAbs in my protein. So I incubated the pAbs with my protein in solution. But when I analysed the complex formation by gel...
15 November 2023 8,480 3 View
Hello everyone, My name is David Vizarraga and I am expressing a single stranded Fv construct. I add a fusion protein like MBP to stabilize the construct and solubilize the protein and this works...
10 March 2023 5,428 0 View
Hi all, my name is David Previously I cloned a protein and everything was perfect. However now I want to clone this same protein but with a PelB sequence in its N-terminal for that I designed the...
02 December 2022 5,572 1 View
Hi all, I want to express differents constructs of Ab such nanobodies or scfv in E. Coli and using pOPIN vectors ( vectors that we are using in our lab). I know there are some complications in...
21 November 2022 2,640 3 View
Hello to everyone, We are study a recombinant protein (Escherichia coli) which has a huge hydrofobic cabity in its structure. By Electron microscopy we saw a extra density in this cavity and we...
21 December 2021 7,701 2 View
Good afternoon my name is David and I am a PhD Student. I want to crystallizate a protein ( It has a small molecular weight, 14kDa) but the problem is the nucleation. The protein forms crystals...
18 April 2017 3,108 6 View
Hi all. After growing and inducting the E.coli cultre I extracted the cells by centrifugation but In the brown pellet there are some black spots. I want to know what is these black spots?. I read...
04 February 2017 3,607 2 View
Hi all. I wrote some times here about a unfold protein topic. The case is I have a pure protein but It is together with DNAk and In the exclusión molecular column I have several peaks that I...
23 January 2017 5,401 4 View
Hi all. I have a pure protein but DNAk is together with my protein. How can I remove DNAk? and If I remove it could the protein suffer any effects? Thank you
17 January 2017 492 3 View
Hi all. I have a pure protein but in SDS-PAGE gel there is DNAk in the solution too ( It is together with my protein). In Superose6 column I have 3 proximate peaks which I cant separated. I think...
17 January 2017 1,713 7 View
Hi all. My protein forms aggregates but I couldnt use any additives like glucose or glycerol because It forms oligomers. Thank you
09 November 2016 9,587 8 View
Hi I would want try EM (negative stain method) with my sample but in size exclusion column I identify 3 different oligomers. Can I do EM? or I need a homogeneous sample. Thank you
10 October 2016 1,242 3 View
I purify my protein but in SDS-PAGE gel I have two proteins ( my protein and DNAk). I cant separate with Size exclusion column so I supposed both proteins are attached. So I want separated my...
26 September 2016 1,026 3 View
Hi all. I try to purify a High molecular weight protein. I explain my case: I did two purification with the same Culture and induction conditions. But when i purified the protein accidentally I...
04 September 2016 1,475 4 View
Hi. I did a lot of trials of protein purification but i couldnt stabilized the protein. I explain the case: When i did the protein purification in the buffer: Tris 50mM NaCl 150mM pH 7,6 the...
30 August 2016 3,775 10 View
Dear all I tried a protein purification with L-Arginine 0,4M but my protein doesnt bind in HisTrap, then i reduced it to 0,2M and in the rest of the buffers I used a 0,4M concentration. With this...
25 August 2016 924 3 View
Hello everyone, I try to purify a High molecular weight protein. But i have a big problem because it forms aggregates. I try to add glycerol (5-10%) or increasing the NaCl concentration of 0,15M...
23 August 2016 4,240 10 View
Hello everyone. I explain my case: I had a protein with this purification protocol: Histrap-MonoQ-Superdex. The protein buffer is Tris 20mM 150mM NaCl pH:7.4. In this case, the protein degraded or...
01 January 1970 4,406 4 View
Hello to everyone. We have a protein which we think it is a metalloprotease, but the experiments that we did, they dont give any clear result. 1) In SDS-PAGE gel, the protein have severals cuts,...
01 January 1970 7,372 5 View
Good afternoon. We are purifying a protein with a 20mM Tris and 150mM NaCl pH:7.4 buffer but when We do the gel filtration column ( HiLoad Superdex 200 16 60) appear two peaks ( 1 at 65ml and the...
01 January 1970 1,272 0 View
Good morning, my name is David. We have a protein structure and now we are looking for epitopes into the structure ( lineal and structural epitopes). When those theorical epitopes are achieved we...
01 January 1970 7,986 3 View
