Ilya Smirnov

6 Questions 24 Answers 0 Followers

Questions related from Ilya Smirnov

Why do antigen concentrations measured in ELISA change depending on sample dilution factor?

Working with home-made sandwich ELISA I found out a strong dependency between measured antigen concentrations and sample dilution factor. I have attached one of such figures. Several blood plasma...

27 September 2017 7,419 23 View

How can I imrove protein synthesis in HEK293 cells?

I need to express a recombinant protein in HEK293 cells. The coding sequnce was cloned into pcDNA6-v5/His A vector without leader peptide. Calcium phosphate or commercial reagent "Turbofect" were...

01 February 2016 3,873 3 View

How can I prevent protein aggregation?

I store recombinant protein (produced in E. coli) in 8 M Urea. It's aliquoted and frozen in -70 deg. Recently I started to notice that certain monoclonal antibodies had stopped recognizing it. I...

03 September 2014 4,237 16 View

Can anyone suggest a good guide for recombinant protein purification from mammalian cell cultures?

I need to purify recombinant proteins from a culture medium using HisTag. I have some experience in purification of bacterial proteins but this task looks rather different and more complex. I...

08 April 2014 301 1 View

What is the reason of protein degradation in 8 M urea?

I purified recombinant protein (expressed in E. coli) 6 months ago and stored it at room temperature in 8 M urea. Previously I observed a thick band of the protein in acrilamide gel. Now the band...

21 March 2014 5,510 24 View

Could anyone help me how to enhance protein expression in stable SP2/0 cells?

Recently I generated stable SP2/0 cells carrying my gene of interest. I used pcDNA6/v5 His A as a vector and blasticidin S as a selective antibiotic. The vector was delivered into cells by...

05 February 2014 766 3 View