RNAse-free filter tips: use despite expiration date that has run out?

I have a lot of RNAse-free filter tips with an expiration date that has run out since a more or less long time (2017 and even 2014!). Do you think I could I use them anyway without much risk to...

11 December 2018 7,255 0 View

Is anyone familiar with denaturation of proteins in native gel?

I have an antibody recognizing my protein in its unfolded state, so I can visualize it by western-blot after SDS-PAGE. Unfortunatly, I can't see it after native-PAGE to follow its...

08 September 2014 492 7 View

How can I avoid HEK cells from forming clumps?

I generated stable HEK lines, but some lines tend to clump (others not), and when I try to separate the cells by pipetting, lots of them dye. As I intend to run flow cytometry, I guess cell...

03 April 2013 7,989 5 View

Are HEK293 Free Style cells suitable for stable selection?

I want to use HEK cell for stable selection, I know 293T are not suitable, and I found a batch of 293F (supposed to be cultured in suspension but that seem to grow very well as adherent cells...)....

01 February 2013 4,888 2 View

HEK293 or 293T for stable line selections with Geneticin?

I want to select stable HEK with Geneticin in order to express a membrane receptor and run functional studies (overexpression is not important). I read that HEK293Ts are not the best cells for...

31 December 2012 2,743 12 View

What is the risk, in terms of stability, to select stable HEK293 lines with a plasmid containing SV40 origin?

My plasmid contains a SV40 origin and a Neo selection marker; if I select 293T cells I have to use another antibiotic (and make a co-transfection with a second plasmid for the selection), if I use...

31 December 2012 2,043 1 View

No title

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

No title

28 February 2021 9,936 2 View

When is it unnecessary to change organisational culture?

As a former management practitioner, I recall a generally accepted view that changing organisational culture is difficult, so you should only attempt it if the organisational change that you are...

27 February 2021 2,595 8 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View