How long doe MCF-7 cells take to attach after thawing and putting in a flask?

I am growing MCF-7 cells and after 4 hours of thawing and putting in a flask, the cells are still floating. I am worried because this is an adherent cell line and should not take this long to...

17 June 2016 4,239 3 View

Is it better to run complex III activity assay without sodium azide?

I am running an assay for activity of mitochondrial complex III. Most protocols use sodium azide / KCN as an inhibitor of complex IV to prevent re oxidation on cytochrome c. However, on addition...

07 March 2016 848 1 View

Do I need to add sodium azide/potassium cyanide and antimycin A while checking for complex 1 enzyme activity?

03 March 2016 382 4 View

Fold change in my end point PCR (checked through a gel) is not the same as array based RT PCR. It is normal or should they match?

I ran an array based q PCR and obtained data for fold change in genes. I followed this up by running end pint PCR followed by a gel. But the fold change in my end point PCR is not the same as as...

27 August 2015 2,126 2 View

Why is there amplification at 52 and 54 degrees Celsius but not at 53 in gradient PCR?

I ran a gradient PCR and there seems to be amplification at 52 and 54 degree celcius but not at 53 degree celcius. What could this possibly mean? My primer and DNA template concentrations were the...

10 April 2015 9,055 5 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

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02 March 2021 3,060 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 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