How to discriminate between active secretion and passive release due to cell death in vitro?

I am about to measure the secretion of a protein into the medium in a cell culture experiment (via exocytosis or extracellular vesicles). However, we have a certain level of death (10-20%) in our...

04 May 2018 3,104 4 View

How is it possible for a hydrophobic sensitive fluorescent probe to show different changes in intensity and blue shifting?

For a fluorescent probe which is sensitive to hydrophobic environment, what factors impact on the intensity and what factors impact the wavelength shifting? In our probe, we observed increase in...

06 July 2017 6,272 13 View

Is there a feasible protocol for the separation of cytoplasmic proteins from cell membrane proteins?

I need a protocol for isolating cytoplasmic content from cell membrane for cell culture cell line.

10 November 2013 2,382 8 View

Drawing error-bar in excel is only based on standard deviation or should we divide it by the square root of iterations?

What do you think, is the drawing error-bar in excel only based on standard deviation or should we divide it by the square root of iterations?

10 November 2013 8,702 4 View

Can someone help in regards to a problem with low efficiency of competent cell preparation?

I am using the CaCl2 method for competent cell preparation but recently I am facing low efficiency of transformation in new competent cells despite several tries. I always test them by doing...

05 June 2013 9,735 17 View

What is the reason for blocking the development of pathology after fibrils injection in a-syn knock out mice?

It seems that endocytosis is the most important factor for expansion of a-syn in brain tissue. Additionally, it seems that knock-out mice have no problem in their essential functions of the brain,...

05 June 2013 1,783 3 View

Can anyone help me to find a protocol for gel extraction without using kits?

I used some kits for gel extraction but I did not get a satisfying result, so I want to do it without using a kit.

04 May 2013 8,838 22 View

How NaOH induce DNA fragmentation in cells, after just 15 min?

We have a DNA fragmentation ELISA kit from Roche. The protocol suggested NaOH treatment for 30 min as a positive control for DNA fragmentation. We did not get significant result, so I become...

04 May 2013 5,315 6 View

How can I treat cells with proteins without using common filters for sterilization?

I have to treat my cells with a-synuclein protein monomers, oligomers and fibrils but for sterilizing we have used 0.22 syringe filter for monomers before fibrillation induction. Now we found that...

04 May 2013 9,778 15 View

Requirement for a software to predict structural changes after a mutation.

I am inducing some mutations in a protein and I want to predict it's structural change. I need a software to figure out this problem. Any suggestions?

03 April 2013 8,307 6 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