Mohd ZIAUDDIN Ansari

12 Questions 17 Answers 0 Followers

Questions related from Mohd ZIAUDDIN Ansari

How can I calculate the amount of Casein Kinase 2 (in mg) required to phosphorylate 1mg of protein having three phosphorylation sites in it?

as one unit of CK2 defined as the amount of required to catalyze the transfer of 1 pmol of phosphate to CK2 Peptide Substrate, RRRADDSDDDDD (100 µM), in 1 minute at 30°C. But I don't know the...

08 August 2015 1,367 2 View

Does dialysing a protein in water effect its structural parameters?

My protein is disordered protein and it does not precipitate in water. However, I am confused regarding dialysis of my protein in water. Should I Dialyize my protein in water and then lyophillize it?

06 June 2015 2,595 7 View

Can I use PMSF and benzamidine both as a protease inhibitor in lysis buffer?

I am purifying a disorder protein (8 kDa) but during binding to Ni-NTA column it got degraded. Currently I am using Benzamidine (2 mM) as a protease inhibitor. can I use PMSF and Benzamidine...

04 April 2015 4,866 3 View

How I can detect pyrophosphorylation in proteins?

I would like to detect pyrophosphorylation in protein. Can you please tell me how I can detect pyrophosphorylation in protein? Is there any antibody available against the pyrophosphate?

01 January 2015 5,297 4 View

Is there any difference in OD between Induced (with IPTG) and uninduced Culture?

Please tell me can i confirm that my protein being expressed by simple checking OD@ 600 of induced and uninduced culture? If yes what will be the difference in OD between induced (with IPTG) and...

12 December 2014 1,966 9 View

Why has the 8 kDa protein cloned in pET21b+ expression vector not expressed ?

My 8 kDa protein cloned in pET21b+ expression vector not showing any expression. Protocol followsed for expression was pET21b+ clone transformed into a E.Coli BL-21 DE3 strain, Induced with 1mM...

12 December 2014 3,164 16 View

Can I reuse Ni-NTA column after using EDTA in elution buffer?

Please suggest if I can reuse Ni-NTA column by recharging it with nickel sulphate, After elution of my protein with EDTA as it stripped Ni from the column. Also suggest me what EDTA conc....

12 December 2014 1,253 9 View

Can anyone help me with the problem in Protein expression and lysis buffer?

I am expressing 6xHis tagged 8.5KDa human Protein (pI~4.19) which cloned into pET21b expression vector and express in BL21 DE3 Codon plus strain. I see expression when I mix bacterial pellet with...

12 December 2014 3,563 13 View

Why do we mostly use only 6xHis Tag to purify Proteins Not 8xHis Or 10xHis?

When ever people make a clone with His Tag to their protein for purification by affinity chromatography why they mostly use only 6 His to protein. What will happened if we put more His suppose 8...

09 September 2014 6,293 13 View

Please suggest me why I am getting very less amount of my protein of interest compare to tagged GST protein after cleavage with Prescission Protease?

I am purifying a 8 KDa protein tagged with GST and cloned in pGEX 6P 2 vector. When I cleaved 8 KDa protein from GST by prescission protease I got very less amount of 8 KDa Protein compare to...

07 July 2014 4,441 13 View

What is the Extinction coefficient of GST?

I am purifying 8 Kda Protein tagged with GST. My 8 Kda protein don't have any tryptophane. So for calculation the concentration of my fusion protein I need extinction coefficient of GST. Please...

07 July 2014 8,842 5 View

Can someone help me with cloning and posttranslational modification of Paramyxoviral membrane protein?

I want to clone a viral membrane protein in pGAPZαA (Yeast expression vector) which have N terminal signal sequence during infection it transport it into host(mammalian cell) golgi body and where...

01 January 2014 8,366 1 View