Protocol for overexpressing NOX4 in a mammalian cell line?

31 January 2024 0 7K Report

I need to overexpress the protein NOX4 in a mammalian cell line. What methods should I use?

List Plasmid suitable for Homology cassette in CRISPR-Cas9 protocol for Knock-In in mammalian cell line ?

Hello, I am working on creating a mutant endothelial cell line having Neomycin knock-in present, I have designed my homology arms of 1kb size having neomycin in between them, I would like to...

25 June 2024 1,197 0 View

Is this mycoplasma contamination?

I had mammalian cancer cell line cultured and fixed for confocal imaging. While observing I found these particles around cell nuclei stained with dapi. I checked the main culture for bacteria but...

15 May 2024 9,102 2 View

Is this mycoplasma contamination?

I had mammalian cancer cells cultuted and fixed on confocal slides. I could see these small small bodies stained with dapi. The main culture flask doesnt show any bacterial growth. Is this...

15 May 2024 6,040 3 View

How do I prevent loss of my H6c7 non-cancerous pancreatic cells during centrifugation?

Everytime I centrifuge my H6c7 cells, the cells don't pellet properly. Though there is a pellet formed, the supernatant has visible particles which I believe to be the cells. I usually centrifuge...

28 April 2024 5,685 1 View

Why does my sample show inconsistent band pattern in western blot?

Hello, I'm having a bit of trouble doing the Western blot experiment. Even though I load the same sample, the band appears inconsistently (in all the trials). (I usually store the samples at -20°C...

18 February 2024 2,863 1 View

Promega Passive Lysis Buffer for Bacterial Cells?

Are there any conditions where it would be possible to lyse gram-negative bacteria using the Promega PLB? I would need to lyse the cells to test for crude enzyme activity but I don’t have a...

30 January 2024 7,625 1 View

Advice for TIDE Primers?

Hi Everyone! I am currently designing primers to validate my CRISPR-KO mammalian cell line using TIDE. The primer is designed about 250bp upstream of the PAM site. Then, Sanger sequencing is...

27 January 2024 2,219 1 View

What is the ideal restriction digestion reaction volume for southern blotting? Can we scale up the volume to 200 ul if gDNA concentration is low?

while working with low concentrated mammalian genomic DNA, can we increase the restriction digestion reaction volume to 200 ul to accommodate the required amount of mammalian genomic DNA for...

22 January 2024 6,952 12 View

How to grow cells in flasks for protein isolation (after treatment with drug) ?

Hey all I want to see the affect of a certain drug on my cell line (cancer cell line). I want to see how it alters the expression of a certain nuclear protein. Since my cells are fast growing...

18 December 2023 4,807 2 View

Need suggestions on bioreactor parameter optimization and PID settings?

Hi, I have established a bioreactor parameters mammalian cell process with the following parameters: Setpoint Deadband PID settings 1) pH-...

14 December 2023 7,651 1 View