How can I eliminate LiCl and BIO (6-Bromoindirubin-3'-oxime) from cell culture conditioned media ?

What are the values needed for gabor filter's parameter for cifar10 data?

Hi, I implemented a code to gabor filter cifar10 data but the images after being filtered and stacked are not clear like the original images. I think the problem is in the way I am using the...

03 March 2021 6,317 1 View

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 View

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02 March 2021 3,060 3 View

How to calculate Total Flavonoid?

I analyzed flavonoid total spectrophotometry, the procedure as below: 2 ml supernatan added 5 ml dd.H2O, then added 0,15 ml NaNO2 5%, 5 minutes later added 0,15 ml AlCl3 10%, After 6 minutes,...

01 March 2021 6,722 1 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

Piyush Gupta

You can either centrifuge it between 5000 to 10,000 rpm for 10 minutes at four degrees or filter it with 0.2 micron syringe filter. Do not use 100% conditioned media. It will kill your cells. You have to titrate it for optimal volume, for example 10% conditioned medium and 90% culture medium.

M.D Pratheesh

Thanks Piyush, This is what we usually does before storing them. But I am not sure whether this high speed centrifugation or 0.2 micro filtration would be enough to remove Licl/ BIO particles from the CM. Please share if you have any literature details here.

What I am thinking right now is to go for a couple of ultracentrifugation steps to isolate exosomes out of it. That also possibly will serve my purpose.