Can anyone share how they label monocytes with CellTracker Orange(CMRA)?

Sharon Lee @Sharon_Lee25

12 December 2017 0 2K Report

I use IMDM as staining buffer (is this ok?),but lose 60% cells after staining.

Also, any idea/comment on why we should avoid amine-containing or thiol-containing buffers as the staining buffer?

Badges
Science method

More Sharon Lee's questions See All

Does company brand of cytokines affect differentiation and/or (M0/M1/M2a) polarization of TH1-P cell line into macrophages?

Concerned that there will be functional differences between products from Peprotech vs. Mitenyi vs. R&D.

05 June 2018 7,386 0 View

What does it mean when transfected macrophages increase in size (forward scatter) and granularity (side scatter)?

Cells treated only the transfection agent or the transfection agent with the miRNA construct (meant for transfection), had a higher forward scatter and side scatter compared to cells that were...

04 May 2018 1,161 1 View

Any suggested tricks or protocols to lift off primary human macrophages from well plates without scraping?

And does scraping activate cells?/ damage surface proteins on cells?

04 May 2018 4,357 0 View

Any comments on whether median fluorescence intensity (MFI) or geometric mean is better to evaluate CD marker expression using flow cytometry?

. .

04 May 2018 6,526 0 View

Any suggestions on how else I can label the nuclei of a cancer cell line that has formed a 3D cellular aggregate?

I used NucBlue Live from ThermoFisher (https://www.thermofisher.com/sg/en/home/references/protocols/cell-and-tissue-analysis/protocols/nucblue-live-readyprobes-protocol.html). I found that this...

04 May 2018 3,880 1 View

Any suggestions on a non-toxic nuclear dye for labelling macrophages while the cells are live?

NucBlue LiveReadyProbes by ThermoFisher (https://www.thermofisher.com/sg/en/home/references/protocols/cell-and-tissue-analysis/protocols/nucblue-live-readyprobes-protocol.html) seemed to seriously...

04 May 2018 5,835 3 View

Is it neccessary to use filtered pipette tips when adding lysis solution to lyse cells for downstream RNA work?

Or is the use of filtered tips only crucial from the point of isolating RNA from the lysate? (i.e. after obtaining the lysate)

03 April 2018 9,465 5 View

Any suggestions on how I can inhibit secretion of cytokines by a cancer cell line without killing the cells?

These 'secretion-inhibited' cancer cells would then be co-cultured together with myeloid cells so I am hoping the environment would not be toxic to both cell types.

03 April 2018 5,179 9 View

What markers can I use to characterize macrophage subsets (M1 and M2)? And what about different monocyte subsets (e.g. patrolling, etc)?

For immunostaining of cells that are fixed.

06 July 2017 7,570 3 View

Comments on the easiest method to attach antibodies to paper for dry storage, where antibodies can be rehydrated when it is required for use?

05 June 2017 5,436 0 View

Do I need specific antibodies to stain for a fusion protein?

I am staining some brain sections stored in cryoprotectant that express a Histone H2B- GFP fusion protein that were generated ~10 years ago. I know I need to enhance signal with an anti-GFP...

07 August 2024 5,338 2 View

AMAXA Lonza nucleofection lower volume of buffer?

Does anyone tried to do nucleofection with AMAXA by Lonza with lower than recommended amount of buffer in the cuvettes (100 ul)?

07 August 2024 4,616 0 View

Why I can't see any band in SDS-PAGE?

Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...

06 August 2024 5,373 2 View

How to determine positive-stained cells in FACS? Use isotype or unstained control?

To compare positive and negative cell populations in flow cytometry, should I compare unstained cells with antibody stained cells? Or with the isotype control? Most papers show comparison with...

06 August 2024 6,728 6 View

Can I multiplex a mouse monoclonal and rat primary for IHC (mouse brain tissue)?

Hi all, I was just wondering if anyone has experience with multiplexing a mouse monoclonal primary and a rat primary. I'm trying to multiplex by incubating them in the same well but was told by a...

06 August 2024 9,710 1 View

Why might the impedance values for DI water and 0.1X PBS buffer solution exhibit a decreasing and increasing trend, respectively over time (HP 4194A)?

Hello everyone, I'm encountering an issue with my electrochemical impedance spectroscopy (EIS) measurements and would appreciate some insights. Experimental Setup: Electrodes: Gold interdigitated...

05 August 2024 3,783 2 View

Does long-term culture of murine Monocytes result in macrophage differentiation?

Hi, I am isolating monocytes from the bone marrow using the Mouse Monocyte EasySep kit. I want to treat these cells and monitor expression of specific markers over the course of 10 days. I will...

04 August 2024 7,282 2 View

Senescence-associated beta galactosidase staining is False Positive in control group?

Hi guys If anyone is currently working on aging cells, you guys would like to give me some advice. I'm testing against biomarker (SA-beta-Gal), I encountered a false positive in the control group...

02 August 2024 6,735 1 View

What is the best blank for nanodrop if I want to read a recombinant protein concentration?

Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC

01 August 2024 967 3 View

Suggestions for the Annexin V Positive Control ?

Hello everyone, I am currently using washed human platelets to stain Annexin V as a procoagulant marker. Additionally, I am staining with PerCP-CD61 to identify platelet cells. So far, I have...

29 July 2024 8,624 1 View