Why does my purified protein have a pellet when I spin it down?

06 July 2015 6,049 8 View

Has anyone made a clonal cell line using C2C12 cells?

If so, how long did it take for single cell to grow and what growth medium composition did you use?

31 December 2014 7,087 1 View

Does anyone have a NIH3T3 monoclonal cell line production protocol?

I am looking for a dilution protocol to make a monoclonal cell line from NIH-3T3 cells. Basically, what I would like to know is: 1.) When you have a single cell isolated what growth...

31 December 2014 2,762 1 View

Do mitochondrial proteins encoded by the nucleus have two bands on western?

06 July 2014 5,547 4 View

What software can I use to predict protein structural changes?

I am looking for a bioinformatic way to predict protein structural changes when I acetylate a specific residue of a protein. Any suggestions?

04 May 2014 8,095 4 View

How many sites have you mutated in a single run of site-directed mutagenesis?

I am trying to introduce 7 point mutations into a 23bp sequence. I have experience doing single point mutations via quikchange site-directed mutagenesis. I was wondering how many point mutations...

04 May 2014 5,435 2 View

What is the most efficient method to screen cells for indels made when using CRISPR/Cas9?

I currently use TOPO cloning to identify cell populations that have indels from CRISPR. I have also used SURVEYOR mismatch assay. Sometimes I can use genotyping PCR and see bands of different...

01 January 1970 5,696 3 View

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02 March 2021 3,060 3 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 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

No title

28 February 2021 9,936 2 View