Haaris Ahsan Safdari

21 Questions 10 Answers 0 Followers

Questions related from Haaris Ahsan Safdari

Publishing in F1000Research??

Hi, Anyone has first hand experience of publishing in F1000Research. I guess it follows open peer review and does not encourage measurable parameters like impact factor! What are pros and cons...

06 June 2019 9,666 1 View

Protein-protein interaction??

Hi, I have two proteins that are his tagged at N-terminus. One of them forms hexamer around 370kDa and other is 17kDa. I want to check their interaction and get Kd value. Which of the following...

04 April 2019 1,556 5 View

Designing mutation experiments??

Hi all, I have a protein with conserved Threonine and one Lysine in the middle of the protein sequence. Being a charged and conserved amino acid across family domain of this protein, I have strong...

03 March 2019 4,925 5 View

E.Coli expression strain for expressing mycobacterial membrane protein gene?

Hi, I have cloned some three different constructs which are putative membrane protein of Mycobacterium tuberculosis into pET28a with N-term His tag. I tried to express them in BL21, Rosetta, C41,...

12 December 2018 1,939 3 View

Bioinformatic tool to predict flexible region in a protein?

Hi, Can anyone please tell some bioinformatics server that helps in predicting the location of flexible region in a protein.?

11 November 2018 6,603 0 View

Citing preprint articles?

Hi, I was just curious to know whether the scientific articles and review published in preprint repository like bioRxiv are citable? If yes, is it good practice to do that since mostly such...

10 October 2018 4,771 4 View

Difference between Coomassie Brilliant Blue R-250 and G-250??

Hi, Can anyone please highlight the difference between Coomassie Brilliant Blue R-250 and G-250?? Which one of them will stain the SDS gel faster?

10 October 2018 2,984 0 View

How to degass a buffer with detergent?

Hi, I want to add about 5 CMC DDM detergent in my Size exclusion buffer. Is it advisable to add detergent and then degass by vacuum or degass the buffer first and then add DDM dropwise slowly??

09 September 2018 4,184 3 View

Publishing in bioRxiv??

Hi, Can someone please tell pros and cons briefly of publishing in preprint servers like bioRxiv?

08 August 2018 7,677 1 View

1M NaOH cleaning overnight?

Hi, One of my colleague put the Superdex 200 10/300 GL column mistakenly in 1M NaOH at flowrate of 0.2ml/min overnight instead of water wash. Will the column be stable? How to know that the...

08 August 2018 5,287 1 View

Zig zag curve in UV detection in akta prime plus?

Hi, I injected my protein in Sephadex 200 column with a flow rate of 0.2 ml/min. The UV baseline is not stable and I am afraid what is happening? Can anyone guide me what is happening? See...

08 August 2018 1,687 0 View

Online repository of Mycobacterium tuberculosis genes in expression vectors of E.coli?

Hi, Does anyone know any website/repository from where I can get mycobacterium tuberculosis gene cloned in E.coli expression vectors on payment basis??

07 July 2018 10,022 0 View

Bioinformatics tools to predict potential ligand?

Hi, Does anyone know any bioinformatics tool or software to predict the potential ligand of a protein?

07 July 2018 8,156 2 View

Rosetta (DE3) plasmid throw out?

Hi, I have rosetta (DE3) with incorporation of tRNAs for AGG, AGA, AUA, CUA, CCC, GGA codons on a compatible chloramphenicol-resistant plasmid. I cloned a gene in kanamycin resistant...

07 July 2018 6,903 2 View

Cleaning of Supherose 6 Increase 30/100 GL?

Hi, I feel that my Supherose 6 column is contaminated with some protein because I get pretty clean SDS gel bands after Ni NTA but after SEC I see multiple bands on SDS PAGE. How should I clean...

04 April 2018 3,045 4 View

Negative stain directly after Ni NTA?

Hi, I am working on a membrane protein (of around 153kDa) which I am eluting by 300mM imidazole (Elution volume-2 ml). I loaded 20ul of my elution fraction on SDS PAGE and I could see my protein...

04 April 2018 5,761 1 View

Need of secondary culture in protein expression?

Hi, Can anyone please explain need of secondary culture in protein expression? Can I directly colony from LB agar plate and grow in LB, then when OD reaches 0.6, Can I induce it with IPTG?

04 April 2018 1,789 0 View

Western blot using anti his antibody?

Hi, Western blot has emerged as confirmatory test for expression of his tagged protein (using anti his antibody). I just want to know that if I perform western blot of elution fraction obtained...

03 March 2018 503 5 View

Concentrating protein by column??

Hi, To concentrate a protein, what speed should be used in centrifugal filter? Does protein stick to membrane? Some people say that concentrating your protein will lead to aggregation, is it true??

03 March 2018 4,108 1 View

NO plasmid after isolation?

Hi, I am cloning a 2.8 kb gene into pet28b vector. I did two ligation reaction. In one of them, I did not put insert (only vector and ligase) (control reaction). Then I transformed 10 microlitre...

02 February 2018 4,391 5 View

When is alkaline phosphatase treatment required in vector?? Is it required every time or only when blunt end cutter enzyme is used?

I am using EcoR1 and Not1 site to cut pET28a plasmid. Do I require alkaline phosphatase treatment before ligation to prevent self religation of vector plasmid?

09 September 2017 790 2 View