Asif Raza

4 Questions 16 Answers 0 Followers

Questions related from Asif Raza

Can anyone advise on a GST purification multiple band problem?

I am trying to purify an insoluble GST-tagged animal membrane protein (70kDa) and I am having problem of eluting multiple (3) bands after column purification. These are having band size of...

12 December 2013 4,571 7 View

Which E. coli strain is best for protein expression?

I want to express a GST fusion protein in pGEX-T2 vector. I have three E. coli strains and I don't know which one is best for the high expression of my fusion protein. I have BL21 De3, Rosetta...

10 October 2013 1,894 5 View

Cell lysis buffer composition for GST tagged protein purification in soluble fraction?

The protein I am working on is quite large (Approx 70kDa, pI-8.96). It is an animal cell membrane protein I am expressing in E.coli. I used the cell lysis buffer (PBS (pH-7.4), PMSF, EDTA and...

10 October 2013 8,039 12 View

Problems with Transformation / DH5-alpha / Plasmid DNA

We have cloned a gene in vector pVITRO2 (GFP-HYG-LacZ; 10KB), and transformed it into DH5-alpha E.coli cells with suitable antibiotic (of course in the right concentration). The problem is, after...

05 May 2013 6,617 15 View