Anil Pooran

8 Questions 18 Answers 0 Followers

Questions related from Anil Pooran

Insoluble protein in insect cell protein expression - any way to increase solubility?

I have been expressing an isoform of a human protein using the baculovirus/insect cell system in sf21 cells. I have been having some problems with expression of this isoform where I have tried 3...

06 June 2013 4,598 7 View

What restriction enzyme site creates a stop codon when it is cut

I'm trying to design a DNA/plasmid construct for eventual protein expression in E. coli. I wanted to know if there is restriction enzyme with a recognition site that, when cut, gives a stop...

03 March 2013 1,247 6 View

Effect of Glutathione S-transferase (GST) and Glutathione on T cell proliferation

I have expressed a human cytokine and two of its splice variants using the insect cell baculovirus system in sf21 cells. I have used a GST fusion protein for one of the mutants as it provided the...

07 July 2012 9,291 2 View

What should the cell viability be at the time of harvest in baculovirus protein expression system?

I am expressing a mutated human cytokine using the baculovirus system in shaker culture using sf21 cells. I have expressed the protein before which has shown up on a gel and western blot. However,...

05 May 2012 4,070 4 View

Gene specific priming strategies in reverse transcription

Hi, I am doing some qPCR measuring gene expression of low copy cytokines in human samples. I am currently doing some optimization for the reverse transcription step (Im doing 2 step RT), more...

01 January 2012 5,255 5 View

Monoclonal antibody generation - what are the best companies supplying these services?

Hi, Has anyone ever used the services of outside companies (in the US or UK) for the generation of custom monoclonal antibodies (using mice) against recombinant proteins (there are no antibodies...

01 January 2012 9,671 5 View

GST recombinant protein does not work in my functional assay

Hi all, I have recently expressed recombinnt IL-4 using insect cell baculovirus system. I have coexpressed in with GST to create an GST-IL-4 fusion protein. Running it on a WB revealed it to be...

07 July 2011 6,855 4 View

Normalizing RTqPCR to total RNA

I wanted some advice on normalizing my qPCR data to total RNA. SO I am doing a 2 step qPCR using absolute quantification and I normalizing first to total RNA then to my HK gene expression. My...

06 June 2011 7,359 5 View