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

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Have someone successfully used the ER alpha (Santa Cruz sc-8002) antibody in immunostaining ?

Santa Cruz now recommends this new antibody (F-10) but I can't find a publication using this reference. I'm working on human et non human primate hypothalamus and so far, it doesn't work. Is...

02 March 2021 10,003 2 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

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 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

Does anyone know an alternative product for cardiac Troponin T antibody for Immunofluorescence (Thermo # MA512960)?

We have been using Thermo Fisher anti-cardiac Troponin T antibody for immunofluorescence staining in FFPE mouse heart sections but it was recently discontinued. The literature shows it to be the...

01 March 2021 4,937 1 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

How does an antibody have full intact antibody + half antibody?

Hello, I was just wondering how does a antibody have full intact antibody + half antibody? Is it because of the drug conjugation?

28 February 2021 1,577 1 View

Does anyone use antinuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples?

I am looking for anti-nuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples. Any suggestions highly appreciated. Thank you.

28 February 2021 6,512 3 View