Will concentrating supernatant (via Amicon filters) give rise to inccurate/biased results in luminex measurements?

11 November 2016 1 2K Report

Especially in the case where supernatant may be low in concentration, whereby supernatant was derived from a low density of cells in culture.

Michael Smalling

Sharon, for accuracy you should get a cell count before lysing or collecting supernatant, then concentration is not a factor if you can correlate it to /per cell.

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

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

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?

11 December 2017 2,469 0 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

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...

09 August 2024 9,621 0 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

How to Add Missing Water Molecules in Protein-Membrane Simulations?

I have protein-membrane simulations (PDB, PSF, DCD) and have noticed that water molecules near the protein are not visible in the simulations. How can I fix this issue? Is there a way to place the...

04 August 2024 1,200 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

Good Day, can I ask for your help on the questionnaire that I want to adopt?

I am currently looking for a questionnaire that relates with my study which is The Effectiveness of Organizational Culture towards employee retention rates

03 August 2024 3,505 6 View