Is using NaOH for lysis of mammalian cells a good idea if you wish to extract a certain compound for LC-MS/MS?

Dear all, good day! I have a question regarding the use of NaOH for lysis of mammalian cells. Is it a common practice? Are there any publications which have used NaOH to perform cell lysis? If...

03 April 2019 5,332 4 View

Which genes or proteins involved in cell migration/motility should i probe for?

Dear all, recently i did a scratch test migration assay on my test compounds incubated with my cell line of interest and it appeared that the cell line seem to exhibit signs of increased cell...

10 November 2018 382 5 View

Should i use serum-free media for MTT assay?

Dear all, recently i am considering if i have been doing my MTT assay wrongly as i have been using my media (with 10% FBS) for my cell viability assay. I realised that the cells were dying at high...

10 November 2018 8,866 2 View

How do you carry out cell migration assay if you want to study if a drug has any effect of increasing cell migration towards another cell type?

Dear all, Recently i have a really crazy and brilliant idea to determine if a compound would be able to increase / decrease the motility of a cell type towards another cell type. How should i go...

09 October 2018 2,941 5 View

Cell death or drug precipitation??

Dear all, recently i found some strange and uncanny sights in the wells of cells treated with my compound of interest. Apparently when i checked the water solubility of the compound of interest,...

09 October 2018 1,427 3 View

Dear all, what are some of the assays i can use to show an increase in intercellular adhesions?

Dear all, i have recently discovered that one of my test compounds is able to promote intercellular aggregation (i.e. the cells are aggregating when treated with the compound). However, i can only...

04 May 2018 7,219 1 View

Dear all, do i require a sterile water control if i used sterile water as the solvent for my drug in cell viability studies?

Dear all, recently i started a cell viability study using presto-blue reagent for the testing. However, i dissolved the drug of interest in sterile water before adding into the wells (96 well...

04 May 2018 8,971 0 View

Dear all, i managed to purchase a Ki67 antibody with a conjugated fluorescent marker. How do i use this to quantify cell proliferation in 3D culture?

As above. Thank you :)

10 November 2017 6,237 0 View

Dear all, i would like to add gelatin and collagen as a liquid solution to my cell culture, but they are solids at room temp. Any suggestions?

As above. Thank you :)

10 November 2017 7,693 4 View

Gene and protein expression woes?

Dear all, Recently I ran qPCR for 3 different sets of treated samples and the trend of the mRNA seemed to exhibit an increasing trend with increase treatment for all the genes tested. However,...

10 November 2017 7,303 4 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