What are the factors taken care of during purification of unstable proteins?

Theoretically, my protein is unstable. till DNA level everything is quite fine, but I think I am losing it during heating with bromophenol for SDS-PAGE. Any suggestion?

03 April 2016 5,136 5 View

How can I interpret double log quenching equation when graph shows exponential change?

03 April 2016 8,497 6 View

Are there any prediction tools for inorganic ligand and solvent interaction?

I am searching an online tool (preferably free of cost) which can theoretically predict interaction (thermodynamic parameters, involving functional groups) of a molecule (in my case its an...

31 December 2014 269 1 View

Is anyone familiar with polyproline tags for purification?

Hello friends, I have a protein (without his-tag) naturally having one pentaproline sequence in the solvent accessible position. Does anybody have experience with purification using the help of...

10 November 2014 1,943 1 View

Why do I observe multiple peaks in flow-through?

I am trying to purify one protein in 2mL agarose-NTA-Ni2+ column at flow rate less than 1mL/min (equilibrated in 100mM phosphate buffer pH 7.4 with 1M NaCl). Multiple peaks can be observed in...

10 November 2014 6,340 10 View

How to prepare FeEDTA?

How to prepare FeEDTA by adding Feso4 7h2o and NaEDTA, avoiding precipitation?

01 February 2014 4,419 3 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

Ester cleavage conditions?

Hi all, I need to cleave the ester in Ethyl 2-amino-1H-imidazole-5-carboxylate to make the carboxylic acid and retrieve the product from solution to use in downstream applications of linking to...

01 March 2021 8,766 3 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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

No title

28 February 2021 9,936 2 View