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

Can anyone suggest a signal peptide for secretion of a recombinant protein of bovine origin?

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

11 December 2013 4,884 1 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

What is the most Operating System (OS) which can be INSTALL on Tiny hard devices like Raspberry Pi (RPi) hardware ?

I'm targeting to deploy a mesh network and manually configure MANET routing protocols. I'm preparing scenarios, architectures, and hard devices needed to do that. Are there some step-by-step...

03 March 2021 1,931 5 View

Which of the Covid-19 Vaccines would elicit the best immunoprotection and why?

I want to know which of the available Covid-19 Vaccines would elicit the best immunological response and why

02 March 2021 1,388 3 View

No title

02 March 2021 3,060 3 View

How can I integrate a routing protocol in a tiny device (e.g. Raspberry Pi)?

Hi Community, I'm facing the issue of integration/compiling a new routing protocol in a WSN simulator. The final goal is to successfully add, configure this routing protocol in hardware devices...

01 March 2021 9,332 6 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Is there a method of determining feed digestibility without the need to re-pellet the feed?

I am aiming to determine the ADC values of a commercial fish feed, but I do not have a steam pelleter or extruder available to me. Ideally, the pellets could be coated with some inert marker....

28 February 2021 7,967 3 View