Which fluorescent dye to use for measuring total ROS production?

Is DCFDA appropriate for measuring total ROS production, or can it only detect certain types of ROS? If I want to measure total ROS production, which dye should I use?

01 March 2021 9,447 3 View

How to calculate Ligand-active site distance in a ligand-protein complex from MD simulation?

I have run a MD simulation of a ligand-protein complex in GROMACS using CHARMM forcefield. Reviewer has asked me to calculate Ligand-activesite distance and binding free energy of ligand with...

27 February 2021 4,703 1 View

How to indicate the degree of litter decomposed by microbes?

Any technique could detect the degree of litter decomposed by microbes? Or any indexes could do this job? I want to split the contribution of microbes during decomposition, so any suggestions...

23 February 2021 1,362 5 View

How to Implement Elastoplasticity in a PCL model in COMSOL Multiphysics?

I am using COMSOL Multiphysics 5.5. I am trying to simulate the tensile test of PCL scaffolds. However, regardless of the strain, I have the elastic region all the time which is not true when it...

23 February 2021 4,050 1 View

Would bisulfide sequencing result be affected if the tissue is fixed by perfusion?

I am looking into DNA methylation of mice as an aging marker along with other assays and recently I added brain IF to the list of assays. For this, I need to fix the brain by transcardial...

09 February 2021 9,012 3 View

Why E1g mode (Molybdenum disulfide) is undetectable in Raman spectroscopy?

I synthesized 1T/2H hybrid phase MoS2 on a graphite foil surface by hydrothermal method, and then analyzed it by Raman spectroscopy. As a result, three new peaks correspond to the 1T phase were...

04 February 2021 6,580 2 View

Distortion problem in Abaqus explicit?

Dear all. I'm modeling the RC column with cyclic displacement by Abaqus-Explicit. The concrete model used Concrete Plastic Damage model. However I'm facing with a error about the excessive...

01 February 2021 7,726 1 View

Can I use 500ng of DNA for restriction digestion?

The recommended amount of DNA for restriction digestion is 1ug in a 50ul rx I've got around 500-600ng of DNA in a total volume of 35ul. Would I get satisfactory results with this amount of DNA for...

31 January 2021 729 6 View

Difficulties purifying Outer Membrane Vesicles using a Sucrose Density Gradient: Unable to detect any protein after this process?

I'm currently trying to purify Outer Membrane Vesicles (OMVs) following their extraction from the bacteria I'm working with. After the initial extraction the concentration of protein seems good...

27 January 2021 6,852 3 View

Where can I find these two papers?

14 January 2021 1,572 3 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

IF troubleshooting: looking at an intracellular protein that localizes to the cell's and how to reduce increase in signal due to cell ruffles?

I am currently trying to stain for PI3KBeta in MDA-MB-231 cells. My current issue is that 1) beta stains all over the cell and 2) there is an increase in signal at the edge of the cell and this...

24 February 2021 8,582 1 View

Looking for advices and tips to improve the staining and the gating strategy in human blood?

Hi, I am staining a human blood with CD19, CD3,CD14 and CD16. My gating strategy in FlowJo goes like this: - First, I gate a general population SSC-A VS FSC-A => Then gate for single cells, =>...

21 February 2021 5,724 4 View

What's the difference between 0.1% triton-X100 and ice-cold methanol in the immunofluorescence assay for intracellular protein staining?

Hi, Guys, I noticed that researchers usually permeabilized the cells with 0.1% Triton-X100 or ice-cold methanol (90%) for an intracellular protein staining, it looks like that the two methods work...

15 February 2021 5,606 4 View

Troubleshooting with immunofluoresence/confocal microscope? What went wrong?

Hi guys, I have no experience and little knowledge on immunofluorescence and confocal microscopy. I tried to visualize the expression of FXR in my cells (THP-1 macrophages), yet I did not...

11 February 2021 562 6 View

How to do immunofluorescence staining of the microtome sections of paraffin-embedded tissue ?

I plan to do immunofluorescence staining of microtome sections of paraffin-embedded metastatic tumor tissue with two or more than two antibodies with different emission spectra. Should I follow...

10 February 2021 3,113 3 View

How do I fix tissue rolling while cutting frozen rat brain section with a cryostat?

I am cutting rat brain sections at 20um at -20C. The tissue is cutting nicely out of the O.C.T solution, but I can not get a decent section to adhere to the slide. I have included an image.

09 February 2021 1,345 5 View

Macrophages in culture not growing/ dying or detaching . Why ?

Hello, I am an undergraduate student doing my dissertation, where I isolate PBMCs from leucocyte cones in order to plate monocytes and allow for macrophage differentiation. Although the numbers...

09 February 2021 5,334 8 View

How can I improve RNA extration from human neutrophils?

I tried several times extrating RNA from human neutrophils and the amount of RNA at the end of the precedure was minimal. I also tried using colunns from QIAGEN. Never had any problem with tissue...

26 January 2021 8,881 4 View

Anti-Src and anti-pY418 Src antibodies for IF?

Hi everyone, Does anyone have a suggestion for anti-Src and anti-pY418 Src antibodies that work well in immunofluorescence on cultured neurons? Thanks a lot!

26 January 2021 866 1 View