Ajeet Singh

9 Questions 24 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 2,652 4 View

Optimum parameters for differential scanning fluorimetry (DSF)?

I am working one bacterial protein pI 4.5, and want to screen optimum stability condition through Differential scanning fluorimetry (DSF). I am new to DSF and don't have much experimental...

26 August 2014 9,553 3 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 6,234 13 View

Phospholipids is not getting soluble during micelles formation?

One of my research protein is bind to phospholipids, for elucidation of this binding mechanism I am trying to make lipid micelle for my interaction study through ITC. I am making stock of...

12 July 2014 8,064 3 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 859 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 9,765 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 3,328 12 View

Competent cell age effects on protein crystal probabilities.

Many people prefer fresh competent cells for protein purification of their crystal structure. Is it really true that using fresh competent cells gives a better chance of getting protein crystals?

05 February 2013 1,978 5 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 1,766 5 View