12 December 2013 1 5K Report

I wish to express a recombinant protein that is not naturally secreted. How to signal a sequence so that it can be secreted out?

More Elamurugan Appavoo's questions See All
How can I make a stable HEK293 cell line using pcDNA3.1?

I wish to make stable HEK293 cells with my gene of interest using pcDNA3.1 How can I proceed?

11 December 2014 8,919 17 View

How do I get good yield of eukaryotically expressed proteins?

i wish to express a protein eukaryotically in good quantity? which system i can go for?  adeno viral vectors are good choice whether these vector have fusion tags to purify? (except yeast...

11 December 2014 9,711 2 View

How do I confirm eukaryotically expressed proteins?

I have cloned my protein into mammalian expression vector pVAC1 and subsequently transfected 6 ug of DNA into 293 cells using lipofectamine...protein will be secreted out of the cell, since vector...

09 October 2014 6,789 7 View

How to add a secretory signal to a protein?

i have a protein that is not naturally secreted out of the cell..so i decided to add a secretory signal (eg. IL-2 signal sequence) at 5' end of the gene... kozak-secretory signal-my gene of...

09 October 2014 1,248 3 View

How does one isolate cytoplasmic and nuclear proteins from cell line (without kits)?

Can anyone suggest a protocol for extraction of nuclear and cytoplasmic proteins from cell line without kits and detergent solution (NP-40)... Can I use Nuclear Extract (NE) buffer (HEPES [20 mM]...

08 September 2014 5,971 7 View

Can anyone suggest how to dissolve TCA precipitated proteins?

I have precipitated cell culture supernatants with TCA (10%) followed by overnight incubation and acetone washing...then I have added Lammeli sample buffer, but I am unable dissolve the protein...

08 September 2014 2,061 17 View

Cloning C-terminal GFP to protein - will this interfere with the expression of GFP?

I have cloned GFP at C-terminal of my protein to confirm its expression by GFP fusion After end of my protein 2 aminoacids are present before start of GFP I wanted to know whether this 2 a.a will...

07 August 2014 3,828 9 View

Transfection of BHK 21 using Lipofectamine LTX plus reagent?

I am doing transfection of BHK 21 cells using Lipofectamine LTX plus. My vector is pVAX1, but I am not getting the expressed protein I included kozak sequence before my gene. The protocol which i...

02 March 2014 4,666 4 View

Isolation of bovine bone-marrow monocytes?

How to isolate monocytes from bone marrow? I need to further culture them for in vitro studies.

11 December 2013 3,458 5 View

Can anyone suggest to me a cell line for in vitro studies on immune systems in bovine animals?

I wish to do in vitro studies to evaluate my protein efficiency to stimulate immune system in relation to bovine animals.

11 December 2013 2,432 4 View

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