Laura Charlier

7 Questions 11 Answers 0 Followers

Questions related from Laura Charlier

How do I improve transfer for low abundance and/or high molecular weight proteins in Western Blots?

I am doing lots of Western Blots and never had any major issues with it. However, I have been struggling to get a nice transfer for a protein of 150 kDa. In the attached figure you see lysates of...

05 April 2025 4,395 3 View

How can I improve RPE1 transfection?

Hi all, I am using FuGENE to transiently transfect RPE1 cells for imaging. I usually seed them on coverslips and transfect them when they have reached 60-70% confluence. For 24-well dishes I use...

03 March 2025 3,366 1 View

Can I store beads coupled to proteins for some days?

I am doing a GST-pulldown. For this, I am coupling GST magnetic agarose beads to my GST fusion proteins; after that I incubate this mixture with my cell lysates. I've made a mistake and started to...

01 February 2025 8,180 3 View

Prioritization of residues in predicted interfaces?

Hi everyone, I'm currently using AI to predict binding interfaces in a protein complex. I used models from AlphaFold2, Rosetta2 and ChaiDiscovery. I analyzed the models with Chimera using the...

12 December 2024 698 1 View

Unwanted primer insertion into template DNA. What went wrong?

Hello, I'm doing site directed mutagenesis. So far, I didn't have any issues. For one construct only, I get insertions that are equal to my primer sequences. Sometimes it is 1, sometimes it's 3...

05 December 2024 7,855 3 View

Poor quality in DNA sequencing results

I sent in my prepped plasmids for DNA sequencing. Unfortunately, the results are very noisy (I attached a screenshot). I double-checked and the samples have the right concentration (100 ng/ul) and...

01 January 1970 6,453 9 View

Biological vs technical replicate in immunofluorescence imaging

Hi all, I am a bit confused with the following situation: I am interested in expression levels within cells upon treatment. I transfected one dish of cells and used a small amount of them for...

01 January 1970 2,974 4 View