How could I stop the autoproteolysis process?

07 August 2017 1,191 6 View

What do you think it is?

03 April 2017 6,929 1 View

A rare purification How could I obtain the monomer or one of the oligomers?

31 December 2016 6,336 2 View

What can i do so my protein will be finished the folding?

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...

31 December 2016 3,167 8 View

Protocol for E.coli expression/induction with a Heat shock step?

Hi all. I want to high the amount of the Heat shock chaperones concentration. I read one option is to do a Heat shock step before the protein induction. Anyone know How could I do? Thank you.

31 December 2016 6,985 2 View

What Could I do for improve on the protein expression for end the fold of my protein ?

31 December 2016 8,390 5 View

Could My protein be affected with the sonication method?

11 December 2016 4,556 12 View

Why after centrifugation at 13,2RPM during 2 hours does my protein broken in the linear monormer?

Hello. My name is David Vizarraga and I am a PhD student. I am trying to purify a protein but I always have a oligomer mix in the solution and I can't crystallize it (because I need a pure...

10 November 2016 5,136 3 View

Which is the perfect buffer for my protein?

Hi all I want to purify a protein (Pi:4,0) and I am use a Typical buffer (50mM Tris pH: 7,4, 200mM NaCl and 10mM glucose ( decreased the aggregates and It is better for the crystallization than...

10 November 2016 9,609 1 View

My protein forms aggregates but I couldnt use additives like glucose or glycerol because It forms oligomers What could I do in my purification step?

10 November 2016 2,739 9 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View