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 8,562 17 View

How do I get good yield of eukaryotically expressed proteins?

11 December 2014 9,665 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 6,749 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 1,200 3 View

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

08 September 2014 5,920 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 2,010 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 3,779 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 4,616 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 3,405 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 2,385 4 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

What kind of fluid could I use for a pitfall trap that also does not invalidate molecular testing?

Hi, I want to start testing pitfall trap to obtain ants samples, but I need to conduct molecular analysis on those insects. So, what kind of fluid can I use? Ethanol expires too early and I need...

03 March 2021 5,978 5 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

No title

02 March 2021 3,060 3 View

How to repeat an operation on an atom selection for multiple times in PyMol?

Hi, I am trying to construct a multi-layer fibril structure from a single layer in PyMol by translating the layer along the fibril axis. For now, I am able to use the Translate command in PyMol...

02 March 2021 4,569 4 View