Non-specific labelling with Click chemistry reaction?

I am working on identification of a small peptide in brain tissue. We have modified the peptide with alkyne modifications for azide-based clicking. For Click reaction we have used 50 uM CuSO4,...

23 September 2019 8,052 3 View

Problem in protein purification?

I have a doudt in adding protease inhibitor. Should i add the cocktail both in cell lysis buffer and column buffer to prevent my protein from degradation during purification and while storage? or...

06 September 2016 4,086 4 View

Can a single amino acid codon changing of a recombinant protein effect protein expression in E.coli?

i have change an amino acid codon asparagine to aspartic acid in my recombinant protein plasmid then transformed in bl21(de3) and used it for expression of my new mutant protein.I expressed it...

19 August 2016 8,032 6 View

What is the minimium possible time to reach OD 0.5 at 600nm by an E.Coli ?

08 August 2016 6,627 10 View

Is there any way I can improve AHL stock preparation for quorum quenching OPA based enzyme assay?

25 January 2016 2,213 3 View

Can someone suggest another scholar besides Hall, that defines hegemony as an ideological force that can produce "the othering"?

In this I mean other profound scholars that have defined hegemony as having the ability to represent those being dominated in a particular regime of representation.

28 January 2015 9,543 6 View

I am looking for scholarly literature on war journalists from developing countries. Can someone help and recommend some work ?

My readings for the past one year has revealed that war journalism is a predominantly western based. But have the developing world indulged in studying their journalists' who cover wars

06 January 2015 9,996 14 View

Can someone help me grasp the concept of memory in journalism?

Beyond Zelizer's work

05 January 2015 1,083 3 View

What is the colour of E coli colonies on EMB agar?

We got purple black, pink and white colonies on EMB agar plate. From that how we will detect E.coli.

10 July 2013 133 41 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

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...

26 February 2021 7,129 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

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View