Manjula Ramu

7 Questions 7 Answers 0 Followers

Questions related from Manjula Ramu

How to prepare Fenton's reagent?

Hi, I want to check the level of Tryptophan oxidation in my protein. For this experiment I want to use fenton's reagent with 0.1mM Fe2+, 0.043mM and 0.05M H2O2. please help in preparing the...

06 December 2017 4,525 4 View

Why sometime agar plates get dried overnight?

Hello everyone, I am facing a problem during transformation. Whenever I use LB agar plates with either kanamycin or amp my plates get dried overnight. Sometime I see colony like dots on plates...

18 January 2016 4,417 4 View

What is the best method to purify basic proteins with pi 7.8?

Hi all, I am purifying a basic protein with pi 7.8. I used 50mM tris 8.8, 300 mm NaCl, 5% glycerol, 0.1mM PMSFand 5mM beta ME in the buffer and performed affinity purification. Eluted with 200mM...

14 April 2015 5,352 5 View

How can I increase the binding affinity of GST-tagged protein to GST sepharose column?

Hi, I am purifying a protein having GST tag. I am using 20mM tris pH 7.5, 200mM NaCl, 5mMDTT and 5% glycerol as the equlibration buffer. Column is of 2ml bed volume (I have hardly 3mg of my...

02 March 2015 7,771 4 View

Why is my protein getting precipitated, even at low concentrations?

I am purifying a recombinant protein (host Rosetta) with his tag. I am getting reasonably good yield (4-5 mg/liter of culture). However after affinity purification using Ni-NTA column, during...

20 June 2014 9,251 4 View

Can anyone help me figure out why I am losing protein during ion exchange chromatography?

I am purifying a protein from a bacterial system and I was able to successfully purify the protein through an affinity column. My protein has a pI of 5.9 and it is stable at higher pH. After...

02 June 2014 8,952 5 View

How much of Dnase can be added and what buffer can be used for cell lysis during protein extraction?

I am isolating protein from more volume of culture. However I am unaware how much dnase to use and also what buffer to use for its activity, if I am dissolving the pellet in 30ml of lysis buffer...

03 July 2013 8,230 4 View