Has anyone got a measurement that could be taken to ensure that an intranasal dose of LPS has been administered successfully?

Dear all, I've been doing some experiments using intranasal dosing of LPS, a technique that has been well described in the literature and I achieve very uniform results (on the whole)....

09 October 2014 4,043 10 View

Anyone know a good IL-1RA antibody for flow cytometry for murine cells?

Hi all, I'm trying to look at IL-1ra expression in lung injury but I'm struggling to find an antibody. Does anyone have any suggestions? Thanks

02 March 2014 9,506 2 View

Does anyone have any experience with measuring AP-1 activation?

I'm looking to study AP-1 activation in some in vivo models of lung injury and was wondering whether anyone has any experience in doing this? Preferably I'd like to study it via flow cytometry but...

01 February 2014 1,197 3 View

Extraction of alveolar macrophages?

Does anyone have any experience with the recovery of alveolar macrophages? We are struggling to get a decent yield because as soon as we infuse saline or media the macrophages seem to adhere very...

10 November 2013 6,905 29 View

Does anybody have any experience assaying Arginase-1 activity in Macrophages?

I'm planning to do some M1 vs M2 macrophage work and am working out the best way to distinguish these subsets. I know you can try this via flow cytometry, however I'm yet to find a suitable Arg-1...

06 July 2013 4,002 16 View

What markers would people recommend when investigating microglial activation by flow cytometry?

I'm currently doing some work with the BV-2 microglial cell line and I'm investigating the effects of a drug on their activation after LPS stimulation. Does anyone have any advice about what some...

04 May 2013 9,836 17 View

Has anybody got any experience with cell cycle synchronisation in either T cells or a microglial cell line?

I can find protocols for HeLa cells, however, we want to synchronise either T cells or monocytes to image them following drug treatment. We're aiming to determine whether the drug causes spindle...

03 April 2013 2,400 20 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