Deekshi Angira

11 Questions 9 Answers 0 Followers

Questions related from Deekshi Angira

Can we apply spherical absorption correction even if the crystal is rectangular in shape ?

We had crystallized an organic compound, collected its SCXRD data at some other place since we do not have in-house data collection facility and solved the crystal structure. The data files that...

06 June 2018 2,400 3 View

How to analyse a CD spectra for a protein containing both alpha and beta secondary structures ?

I recently performed CD analysis for my protein of interest in 50mM sodium phosphate buffer. The curve that I got had a dip at 218 nm and 208 nm with a peak at 195 nm ( Keeping in mind that alpha...

10 October 2016 637 6 View

Can anyone explain PLAT340 Low Bond Precision on C-C Bonds and PLAT213 ADP max/min ratio error ? What is 'U' here in this error?

I recently solved a crystal structure for small molecule and submitted the cif file for scanning alerts, if any. I was able to rectify all the alerts but got stuck with this one. PLAT213 Type_2...

08 August 2016 1,707 1 View

Can we use linear DNA fragment as a template for PCR ? If yes, then what should be the minimum number of base pairs in that fragment ?

I am curious to know that in case we plan to add a ~25 base pairs long site in our vector , how can we do that. We do not have any template for the given site so I was wondering whether we can get...

06 June 2016 3,751 3 View

How to determine the concentration of substrate / inhibitor to be used with protein for co-crystallization ?

I am planning to set up a co-crystallization trial for my protein. I am confused as to what parameters should we consider in terms Km or Ki in order to decide an optimum concentration of...

02 February 2016 8,502 4 View

Any advice on why there are multiple bands even after affinity chromatography and FPLC?

I am trying to purify a protein ( Kinase domain) , ~80 kD in size. I loaded the crude extract after expression on affinity column ( Ni-NTA) . On running SDS gel I got multiple contaminating bands...

09 September 2015 1,596 7 View

Is it possible that a tagged protein degrades and separates from the tag during protein expression?

My protein (16 kD) is tagged with Maltose binding protein ( 42.5 kD) at N terminal and Histidine tag( ~1 kD) at C terminal. When I am expressing my protein in Rosetta Gami pLysS and BL21DE3, after...

06 June 2015 5,456 8 View

Is it possible for a protein to form dimer when expressed in bacteria ?

The protein that I am working on is a 16 kD protein. When I am trying to express this protein in E. coli and performing SDS PAGE I am getting a very faint band at 16 kD but at the same time I am...

04 April 2015 9,576 10 View

Is it possible that a very low amount of protein is expressed in uninduced culture too ?

I induced a culture (Rosetta gami pLysS) with IPTG ( 0.5 mM and 1 mM) at 37 degrees. Together with it I also kept a control ( uninduced). After lysis, when I run SDS-PAGE gel, I get a band at...

01 January 2015 7,057 5 View

Is there any possible method to prevent our protein from going into inclusion bodies after expression, apart from using solubility tags?

The protein that I am working on has a tendency to form inclusion bodies. So, I want to know that there is anything that we can add or any precaution that we can take during protein expression...

01 January 2015 1,682 13 View

BL21 DE3, BL21 DE3 pLysS and BL21 DE3 pLys E : Preferred choice ??

Out of these three, which strain should be preferred for expression of protein that has a tendency to form inclusion bodies?

09 September 2014 6,094 15 View