I am working on the megakaryocyte development. I need this cell line for my study.
CMK seems to be only distributed through DSMZ. See:
http://web.expasy.org/cellosaurus/CVCL_0216
https://www.dsmz.de/catalogues/details/culture/ACC-392.html
To validate a miRNA target mRNA can we do it in vitro by adding everything in a test tube? If not, what are the difficulties and how can we overcome this?
08 September 2014 981 4 View
I am using realtime PCR approach to measure mtDNA copy number with mitochondrial tRNA or rRNA and nuclear B2M primers. I isolated DNA from both the samples control and treated then I did real...
06 July 2014 7,645 8 View
When I perform an MTT assay after adding DMSO, only a brown colour is appearing. I am working with suspension cells and I want to know the cytotoxicity of a compound. After 24 hours of seeding...
11 December 2013 7,328 15 View
I want to check the miRNA levels and miRNA transcript is short in length. How to design primer for a specific mature miRNA?
11 December 2013 9,940 7 View
I want to stain the mitochondria in suspension cells by using mitotracker red (Invitrogen). Can anybody explain protocol to observe them in fluorescence microscope?
11 December 2013 829 12 View
Buffer composition is 50mM tris pH7.5(p.I 6) 150mM Nacl (tried upto 300mM) 5mM BME 1mM pmsf 10% glycerol. Even it is also aggregating after NI -NTA purification but I am diluting protein then it...
11 December 2013 9,452 6 View
If I want to design primers or mature form of a miRNA it is very short, so how to get primers. Can anybody explain in detail?
08 September 2013 937 2 View
When I am performing an experiment I could not observe the any signal. Usually I use FITC tagged antibody but I am suspecting that I have problem with sample preparation and cells may be damaging.
08 September 2013 8,117 14 View
Can anyone send me a detailed procedure of miRNA cloning in a mammalian expression vector?
11 December 2012 659 0 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
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
I would like to perform transfection with the reagent DharmaFECT Duo (Horizon) on the AGS cell line. Could you please inform me of the optimal concentration to use without causing cytotoxicity in...
03 August 2024 3,851 1 View
Hello, if you made a transwell assay where you incubate the cells with a nanoparticle-encapsulated drug and considering that in the oposite compartment you'll have both the free and encapsulte...
03 August 2024 2,730 1 View