In flow cytometry single histogram of a single phosphoprotein splitted into two peaks. Even that is a stable cancer cell line. Why is this?
I have purified an antibody the final concentration is 0.42mg/ml. i want to titrate antibody, so that i can pick up 0.5 microgram/ml to 5microgram/ml concentration in 100 microL staining medium. i...
31 December 2014 2,652 2 View
Can anyone tell me how to save the cells (Human Leukemic cell lines) after treating with drugs before preparing proteins for Mass spectra? i have other things to do so i have planned to save the...
11 December 2014 6,353 2 View
Hi guys. Can anyone tell me the suitable stimulation conditions to stimulate pStat1, pStat3, pStat4, p38 and pAkt. I am working with mouse spleen, bone marrow and murine cell line BaF3. I am...
10 November 2014 8,197 2 View
I performed multicolour analysis in flow cytometry. I stained for FITC, PercpCy5, PercpCy7, PE and PB on cancer cells. I compensated using Beads and used FMos for gating population, but when I run...
03 April 2014 4,150 14 View
I am working on phosphoflow for specific proteins. In the histogram sometimes a single histogram split into two peaks for single phosphoprotein, but sometime for the same protein that peak...
11 December 2013 5,684 5 View
I added PI to the cell line that I am interested to exclude dead cells. Then run the experiment in FACS canto. But GFP is sowing the peak at 10 to the power of 5 in FITC channel for all the...
11 December 2013 5,469 5 View
Which channel should I use to see the population of GFP? Can I use the APC channel?
11 December 2013 9,203 9 View
I am working on phosphoflow in FACS and have stained the cells with surface markers and intercellular markers. I got the FSC and SCC results and histogram but I am confused about how to interpret...
10 November 2013 10,032 4 View
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
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
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
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
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
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
Women, on the other hand, can become physically aroused (increased blood flow in the reproductive organs) without becoming psychologically aroused even in the slightest. (Robert Weiss)
05 August 2024 9,537 2 View
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
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
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