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Questions related from Santhosh Gatreddi
I have to calculate the concentration of free divalent metal ion that is available in a enzyme reaction based on the log K value of particular metal-ATP complex. Because the reaction also contains...
30 July 2017 2,627 1 View
I have obtained a graph which was plotted by using the equation v= kcat[S]/(Km + [S] + [S2]/Kis) . Substrate concentration varied from 0.1-5mM. But when iam going for data points lower than...
02 May 2017 6,561 6 View
Iam measuring the absorption of substrate in a enzymatic reaction to find out the kinetic parameters Km and Vmax. Plot of the curve (absorbance vs time) obtained in a exponential decay manner. To...
05 December 2016 7,449 3 View
Iam trying to prepare 0.1M ZnCl2 solution but it is forming white precipitate. What are factors that influence its solubility in water or what might be the probable solvent to make it soluble.
05 May 2015 1,360 10 View
What is the critical concentration of a ligand that is used to cocrystallize a protein along with its ligand and how the varying concentration of the ligand effects the crystallization.
02 May 2015 9,355 6 View
I have purified a protein of 37.5 kDa by Ni-NTA and finally got 9.2mg/ml of protein after gel filtration (monomeric). After concentration we have checked the homogeneity of the concentrated...
15 January 2015 200 20 View
I have done enzyme assay for my purified protein, according to the previously reported protocol one proton will be released per reaction and it was monitored by change in the pH of the reaction...
07 December 2014 7,785 15 View
I have to express a series of proteins with different molecular weights ranging from 20-36 kDa. I need to express these proteins with pET vectors in E.coli and according to some research articles...
07 August 2014 3,705 5 View
I am working on a protein from Leishmania donovani DD8 strain but according to gebeDB genome sequence of Leishmania donovani BPK282A1 strain was available. If we have crystallized the protein from...
01 August 2014 3,022 7 View
I have solubilized my protein with 0.3% sarcosine and purified by using Ni-NTA,during purification most protein is going into flow through. I have diluted my sonicated sample to 0.1% sarcosine...
17 May 2014 9,102 6 View
How will metals be recruited in proper coordination and how will a particular metal be selected to its target protein? If I want to express a metalloprotein that needs two ions for its stability...
28 February 2014 351 4 View
To know the reason I kept separate reactions each with 1ug of plasmid and the reactions as follows. 1.Plasmid with nuclease free water 2.Plasmid+water+buffer Incubated at 37c for 4hrs and rsults...
21 February 2014 9,449 4 View
Which parameters can affect its shape during crystallization?
28 October 2013 240 8 View
It was a 1% agarose gel and run at 90 volts
14 October 2013 1,060 0 View
If not, which is the better concentrator that is suitable for the exclusion of non-specific bands (at 20kDa)?
24 September 2013 743 12 View
I have read some articles, in which they have used BL21 and in other some cases it is M15 cells. So, which is the better host strain for expression and why ?
16 September 2013 5,550 10 View
As most of the proteins are going into the insoluble fraction as an aggregate and a very low amount in soluble fraction...
16 September 2013 5,150 5 View
I have taken my sonicated sample and kept it for centrifugation (10000rpm for 15min) to separate the pellet from supernatant. After that, I centrifuged my supernatant @15000rpm for 30 min and...
07 May 2013 5,998 36 View
How does the isoelectric pH of a protein effect its solubility during its purification from bacteria?
06 May 2013 5,917 2 View
