Ajeet Singh

6 Questions 20 Answers 0 Followers

Questions related from Ajeet Singh

What is the role of buffer pH and salt conc. in crystallization?

I am working on bacterial protein, which will be crystallized for structural study. My protein MW is 22kDa and pI is 9.2. I have tried crystallization trial in 20mM Tris, 150mM KCl and 5 %...

25 November 2014 2,644 4 View

What are optimum conc. of salt in protein crystallization?

I am working on bacterial protein crystallization (protein pI 9.2). During purification, I usually maintain 200mM KCl salt (20 mM Tris 7.5, 10% glycerol) in final elution buffer and followed by...

16 August 2014 6,224 13 View

How to make a protein soluble?

My expressed protein is being collected in the pellet fraction and I have used different strategies for making it soluble but still its coming in the pellet. I have used strategies like...

11 February 2014 850 31 View

What to do when protein is getting precipitated during dialysis?

I purified my recombinant His tag protein in Tris, pH 7.5 (pI of protein 8.9) using 400mM Imidazole conc. in elution buffer. I dialysed my protein for removing imidazole but it is being...

04 October 2013 9,754 21 View

Non specific amplification in PCR?

I am getting non specific PCR, I have tried with increase temperature and decreasing primer concentration but still non specificity is there? What parameters should I consider now?

01 October 2013 3,318 12 View

Which buffer is the better choice for membrane protein purification.

I have tried to purify membrane protein in HEPES buffer but still there are some other buffers there. Which buffer and other biochemical parameters should be followed for better protein induction...

16 January 2013 1,756 5 View