Philippe Paget-Bailly

13 Questions 44 Answers 0 Followers

Questions related from Philippe Paget-Bailly

What tag to use inside the sequence of an hydrophobic transmembrane protein ?

Hi everyone, I want to study the translation efficiency of multiple version of the same protein coded by different genotypes. To do so I need to conserve the translation initiation context ->...

30 April 2024 6,658 1 View

What is the shortest sequence encoding a fluorescent protein ?

Hi everyone, everything is in the title question ! To the best of your knowledge, what is the shortest sequence you know that translate into a fluorescent protein that could be used for flow...

08 March 2024 1,732 1 View

Why the concentration of my expression vector drop with time and how to stop it ?

Hello everyone, I use several pcDNA3.1 expression vectors to transfect cells. The vectors were prepared by midi-prep a year ago and diluted in TE buffer. Now that I run new experiments, I...

05 March 2024 1,841 3 View

Who has experience with transfecting HaCaT keratinocytes ?

Hi everyone, I'm currently working on the HaCaT keratinocyte cell line. I have them in culture under low and high calcium concentration to keep them undifferenciated or differenciated,...

05 March 2024 2,120 0 View

How to get rid of chelex resin before 0.22µm filtration ?

Hello everyone, I'm curently working on HaCaT cells which require low calcium medium to return to undifferentiate state. To do so, I need to chelate my FBS for 1h with chelex resin among other...

13 December 2023 4,945 2 View

Does 5'UTR length can influence alternative splicing ?

Hello every one, I'm working on the in vitro expression regulation of a viral gene. I'm looking at the splicing efficiency of several versions of the gene (comming from different genotypes). To...

11 September 2023 9,657 1 View

Would you harvest cells in medium with or without FCS ?

Hello everyone, I came across a question while harvesting my U2OS transfected cells. After trypsination, I resuspend my cells in cold medium with 10% FCS and I centrifuge them into four...

14 June 2023 1,687 5 View

Why my deltaATG mRNA is in polysome fractions ?

Hello everyone, I'm doing polysome profiling to answer a specific question. I study a gene which produce an alternative transcript and it's translation is controversial (extremely lucky if you...

17 December 2020 7,770 2 View

What could explain the presence of deltaATG transcript in polysome fraction ?

Hello everyone, I'm doing polysome profiling to answer a specific question. I study a gene which produce an alternative transcript and it's translation is controversial (extremely lucky if you...

24 November 2020 4,175 0 View

Does polysome profiling in 4mL tube instead of 12 mL needs a scale down in ultracentrifugation time ?

Hello everyone, I recently started several polysome profiling experiment and something bugs me out. Every protocols I found use 12mL tube. They load their lysate containing ribosomes attached to...

20 September 2020 8,870 10 View

What is the best Amplex red peroxide assay protocol ?

Hi everyone, I want to use the amplex red assay to quantify H2O2 levels in different cellular clones. Many protocols I found online or previous answers on RG contradict each other. The major...

13 April 2018 8,963 4 View

What is the effect of a tag on a small protein ?

Hi everyone, here's my problem, I'm working on the potential role in carcinogenesis of a small protein of 50 aa (E6*I from Human Papillomavirus type 16). Many cellular effects caused by this...

22 September 2017 8,596 2 View

On the need of switching our -80°C storage to -70°C for environmental reasons.

Hi everyone, we recently switched most of our -80°C freezers to the absolutely scorching -70°C in my lab. We did so because apparently the -80°C guideline originated from a technical limit...

01 January 1970 5,994 3 View