I'm using puromycin for selection, usually I'm making 50 ml of growth media that lasts for about a week, I'm wondering if Its possible to make 500ml and have the puro remain stable for a whole month.
I'm interested in the entry mechanism of the Zika Virus into U937 Monocytes. To the best of my knowledge, the proteins responsible for Zika entry are DC-SIGN, AXL, Tyro3, Mer, TIM and...
17 February 2019 8,523 3 View
i need to ask that when I transduce my HEK293T cells with Lentivirus, how much time will they require for transduction because when I add lentivirus and then change the media after 24 hours, my...
07 July 2024 2,946 1 View
Hello, everyone. I would like ask one question. I transfected fibroblasts with EF1A-myod-puro using lipofectamine 3000. Promoter for puro is CMV. After transfection 48 hours, I did puromycin...
23 June 2024 4,902 3 View
I am looking at puromycin incorporation rate by flow cytometry using a specific anti-puro-antibody which I use to stain different cell types. For some cells the signal is high (i.e. Puro MFI),...
12 June 2024 1,110 1 View
My transduced cell can resist Puromycin and 99% express GFP, but it cannot show me the difference in the knockdown. What could be the possible reason?
31 May 2024 454 2 View
Hello all, I want to immortalize human adult fibroblasts with a lentivirus plasmid containing hTERT under the promoter EF1A and puromycin resistance under the promotor mPGK. I performed a...
14 May 2024 6,842 3 View
Hello dear researchers,i have transduced my HEK293T cells with IL-2 and then puromycin screening was done, the cells were showing good egfp flourescence ,but then due to some reasons I have to...
14 April 2024 7,285 2 View
I have been trying to make a stable cell line expressing my GOI using lentiviral transduction of HEK293T cells. To do so I ordered a lentiviral transfer plasmid from addgene which had a gene in...
26 March 2024 7,333 2 View
I am trying to express m6A protein and a fluorescence one RFP in the piggyback vector. I was thinking first of using the P2A to separate between the m6A and the RFP protein but then I was afraid...
12 March 2024 9,713 0 View
I have knocked down my gene of interest by lentiviral transduction. I revived the vials and continue growing cells for 72 h without any selection. After confluency, I passaged the cells later 24...
28 February 2024 4,032 1 View
Hi all. I performed CRISPR knockout by lentiviral transduction. Cells were grown in puromycin for a period of time, protein was extracted and cells were also frozen down. A western blot showed...
24 February 2024 1,550 1 View