Does anyone have a protocol for protein extraction after exosome isolation by UC or Exoquick for an SDS-PAGE separation and Mass-Spec analysis?

Can you connect an HPLC to a Mass Spec only at a certain time point?

Can anyone explain this method? Especially the last statement where it says only at 1.5 to 2.5mins was the MS/MS connected to the UPLC. How is that possible, is it a feature in this specific...

11 August 2024 8,141 3 View

GC-MS retention index prediticon?

Hello experts, Does anyone know any free software about retention index prediction ?

08 August 2024 7,403 2 View

How to calculate CCS for Sodiated adduct ions and Multiply Charged Ions?

I'm currently working on calculating the collision cross section (CCS) for various ions, and I'm facing challenges when dealing with sodiated and multiply charged ions. Most of the resources I’ve...

08 August 2024 8,329 0 View

Where to find a gene list for CRISPRa/i library screening of regulatory factors that affect pathogenic Th17 differentiation in PBMC?

I want to perform a CRISPRa/i library screening for candidate regulatory factors that affect pathogenic Th17 differentiation in PBMC. But I am wondering how to define the size of gene library, so...

31 July 2024 2,150 0 View

Hello dear researchers. I would like to know what is a normalized mass in an EDX analysis?

EDX analisys

31 July 2024 5,947 2 View

Gas chromatography RT detection?

I am working on algal extract to which gas chromatography (Not GC-MS) spectrum I want to discover. My question is can we identify specific compounds using retention time if I compared the RT with...

29 July 2024 8,034 4 View

MAGeCK Plotting Error: Logarithmic Axis Must Have Positive Limits?

I have been running a MAGeCK test command on the terminal for a CRISPR screen to rank sgRNAs and genes based on the read count tables. However, I get only the plot of the top-ranked genes...

28 July 2024 9,811 1 View

Screening of EPS producer LAB?

if you have MRS broth already prepared, what is the manner you will follow for preparing hyper-sweet medium for screening of EPS producer LAB? what the concentration of saccharose or glucose you...

26 July 2024 6,606 0 View

I found the zeta potential of exosomes in DLS and it was -8mv. So is it correct or not ?

What should be the zeta potential of bovine milk exosomes

25 July 2024 7,376 5 View

Can I use sonicator only to lyse the exosomes and measure their protein contents using BCA assay kit?

Hello, I want to ask regarding exosome protein quantification method. Is it necessary to lyse exosomes before measuring their protein contents using BCA assay kit? Or can I just straight away...

23 July 2024 4,603 4 View

Jeroen van bergenhenegouwen

its important that you mention what the source of your material is. The answer to your question will for a large part be determined whether your exosomes are isolated from serum/supernatants/ex-vivo/etc etc. for example, the isolation of exosome from milk or serum can be very tedious due to the complexity of the matrix. This might be helpfull: http://www.ncbi.nlm.nih.gov/pubmed/24009894; http://www.ncbi.nlm.nih.gov/pubmed/22722367

Shlomit Kenigsberg

Thank you for your answer,I am isolating exosomes from serum and other body fluids and we found that even after extensive UC and sucrose gradient there are still proteins carry-over. also in most publication with Mass-spec analysis, you can not locate any tetraspanins.

If you currently do bottom-up sucrose gradient..try top-down..i have heard that this really improves the quality of the EV fraction.

Kazem Abdian

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