How transfectable are U87MG cells?

I was wondering if someone might have experience transfecting expression plasmids into U87MG (glioblastoma) cells? I would like to transfect in an expression plasmid and overexpress a protein in...

05 June 2018 389 1 View

How "transfectable" are mouse embryo fibroblasts (MEFs)?

I have some +/+ and -/- MEFs of a certain gene and want to add back in WT and various mutant cDNA of this gene, to determine functionality. I was planning to transfect the various cDNA's/plasmids...

04 May 2017 5,985 3 View

Has anyone used a colorometric or fluorometric kit to measure free fatty acids in cell culture cells?

I want to measure possible changes in fatty acids in various cancer cells in culture after modulating the expression of a certain protein? I don't have the $ or the resources to do this...

04 May 2016 628 1 View

Is there an upper limit to the size of a protein that can be expressed in BL-21 bacteria?

I understand every protein is different regarding induction and solubility. I'm just wondering if there is a rule of thumb regarding a molecular weight limit. I'm thinking about trying to express...

11 December 2014 1,394 7 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

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02 March 2021 3,060 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

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28 February 2021 9,936 2 View

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 View