Why is there no protein expression but antibiotic resistance in transduced A549 cells with pLVX-TetOne-Puro lentivirus?

Hi, I have been trying to transduce A549 and Calu-3 cells using the Lenti-X Tet-One inducible expression system (TakaraBio). My gene of interest is inserted in a pLVX-TetOne-Puro plasmid and has...

15 February 2021 8,462 3 View

Any recommendations for tetR antibody for IHC?

I am planning an experiment where I've to detect cells which are expressing TetR, my vector is so big that I can't have a reporter in it. I was thinking if I can detect the expression of TetR...

24 September 2020 1,041 4 View

Should I add penicillin/streptomycin into cell culture media for doxycycline inducible system?

Hello, since I have no experience in Tet-on system in drosophila S2 cell culture, I need some technical help... Should I add P/S into the cell culture media? What's the doxycycline concentration...

17 September 2020 7,497 5 View

Why is doxycycline drastically affecting my control-vector cell line?

I am using a Tet-On system to observe how over-expression of a certain gene affects cell proliferation. I have my cell line with my gene of interest and an EGFP control line. Using western blot I...

28 July 2020 7,127 2 View

Do we need further research on HCQ and Azithromycin combination ?

Both HCQ and Azithromycin are known cardiotoxic drugs. Provided that COVID-19 infection also leads to myocardial injury and may present with variety of cardiovascular conditions. Will this...

15 July 2020 5,061 8 View

Can the pcDNA5FRT/TO vector be used for transient overexpression of a gene in a common cell line?

01 March 2020 3,206 2 View

Problem with inducing knockdown using pTRIPZ vector?

I recently began using the pTRIPZ system. I added my specific shRNA to knock down my gene of interest, and then packaged the vector into a lentivirus for transduction. After transduction, I...

07 August 2019 9,195 0 View

What is the optimum time required for treating transfected HeLa cells with Doxycycline to induce the expression of recombinant GFP tagged protein?

Hi all, I am going to perform immunofluorescence analysis of transfected HeLa cells to determine the localization of the protein of interest.The protein is tagged with GFP and I am using...

05 June 2019 2,230 2 View

Does anyone have any experience in generating C2C12 and ST2 stable cell lines using lentiviral vectors?

I am trying to stably express either a WT or a mutant form of my protein of interest under the control of doxycycline (using pTRIPZ vector) in C2C12 and ST2 cells to study it's role in bone...

04 May 2019 1,131 2 View

Doxycycline hydrochloride or doxycycline hyclate for gene expression induction: any difference?

Is there any difference in using doxycycline hydrochloride or doxycycline hyclate salt for doxycycline-inducible gene expression?Any special requirements for doxycycline reagents used for gene...

31 December 2018 1,351 3 View