How to increase the pH tolerance of an Enzyme ?

There is an enzyme which pH optima is 6.5, we want to make this enzyme working well at pH 4.5, without disturbing its catalytic efficiency for a reason of industrial fermentation process. The...

06 July 2018 8,909 6 View

Is it possible that Km of key residue mutant is similar to wild type enzyme ?

I am getting Km value of a mutant which is similar to its wild type. How it is possible ? The residue is quite important for substrate binding. Is it possible ? please suggest !

10 November 2017 5,476 4 View

Is it fine to mutate some conserved residues in the active site ?

I am planning to make the wild type enzyme pH tolerant (acidic) without affecting the activity of the native enzyme. Here i am thinking to introduce some residues like lysine or ariginine in place...

08 September 2017 1,322 5 View

Which tool is best for showing secondary structure element on the sequence alignment other than ESPRIPT (which can use the pdb file to do it) ?

I am trying to find some tool or server which put secondary structure elements on top of the sequence alignment which ESPRIPT does very well. But is there any other tool which I can use to show...

02 March 2015 712 5 View

Why the protein after Site Directed Mutagensis is not expressed ?

I have done whole plasmid amplication using XT-20 high fidelity enzyme from merck. After induction with 0.3mM IPTG i did not see any expression of the protein.I am suspecting some mutation has...

01 February 2015 3,091 6 View

How can I isolate double stranded DNA from single stranded in a mixture?

09 October 2014 6,666 5 View

Why is the Difference between Rwork and Rfree 10% when refined at 3.2 Angstrom?

I am little bothered that my structure refined at 3.2 Angstrom, have a difference of 10 % without and with solvent contents. The difference is not changing. completeness is 97.6 % at 3.2 Angstrom....

06 July 2014 6,790 8 View

How does one shade the conserved and non-conserved protein sequences after MSA?

I have used a boxshade server, but here I am finding difficulties as it puts sequence numbers here and there after the job. So does anybody have an idea to do the same without any erroneous job of...

06 July 2014 9,086 4 View

What is "Refmac sigma-AA weighted map data"?

06 July 2014 7,131 5 View

How to improve the quality of my crystal?

As I have purified my protein and set crystallization screen and I tried various way to optimize the crystal quality, such as seeding, changing pH, PEG concentration, salt concentration and...

04 May 2014 8,551 10 View

What is the most Operating System (OS) which can be INSTALL on Tiny hard devices like Raspberry Pi (RPi) hardware ?

I'm targeting to deploy a mesh network and manually configure MANET routing protocols. I'm preparing scenarios, architectures, and hard devices needed to do that. Are there some step-by-step...

03 March 2021 1,931 5 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

How can I integrate a routing protocol in a tiny device (e.g. Raspberry Pi)?

Hi Community, I'm facing the issue of integration/compiling a new routing protocol in a WSN simulator. The final goal is to successfully add, configure this routing protocol in hardware devices...

01 March 2021 9,332 6 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