Does anyone have any recommendation for microscopy-based assay for T-cell lytic function?

We are working with primary human T cells and are searching for a good assay to look at their lytic function. The chromium release assay is an option, but we're looking to avoid radioactivity...

07 August 2016 407 1 View

Does anyone have any advice for running primary human monocytes on a western blot?

19 July 2016 1,036 5 View

Does anyone have any advice for running primary human monocytes on a western blot?

06 July 2016 9,137 6 View

How is the chamber removed in BD Biocoat 8-well cell culture slides?

Has anyone used BD Biocoat 8-well cell culture chambers before? If so do you have any advice on how to remove the chamber without soaking in an alcohol? I cannot permeabilize the cells I am...

10 November 2014 7,816 5 View

How is the chamber removed in BD Biocoat 8-well cell culture slides?

Has anyone used BD Biocoat 8-well cell culture chambers before? If so do you have any advice on how to remove the chamber without soaking in an alcohol? I cannot permeabilize the cells I am...

03 November 2014 5,807 4 View

Does anyone have any advice on quantification of mitochondrial probes using FACS vs. microscopy?

We have 2 conditions where the morphology and signal of several mitochondrial probes are different. I keep coming across caveats associated with both techniques when it comes to quantification....

03 April 2014 4,421 0 View

What is a good protocol to dry cells that have been seeded on slides for immunofluorescence?

I have come across a few protocols using different increments of ethanol, but most of these refer to tissue sections. Most others in our lab have also done this with tissue sections. I am using a...

11 December 2013 521 5 View

Plasma membrane proteins and western blot

Is anyone familiar with plasma membrane protein aggregation in samples for western blot? I am getting good signal but the pattern makes no sense! I did some brief reading and it seems plasma...

26 October 2013 7,105 19 View

Plasma membrane proteins and western blot

Is anyone familiar with plasma membrane protein aggregation in samples for western blot? I am getting good signal but the pattern makes no sense! I did some brief reading and it seems plasma...

09 October 2013 6,587 13 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

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