6 Is your exosome prep contaminated with IgG and/or beads?
A: We use specific reagents to grab exosomes. We did not use antibody or beads for the isolation. So the last word exosome prep simply is not contaminated with artificial IgG or beads.

7, Do your exosome prep incorporates Albumin?
A: Some sorts of cells akin to hepatocytes carries enriched Albumin, and these liver derived exosomes will likely be launched into the circulation system. So, if the exosomes from these kinds of samples are isolated, it would incorporates Albumin.

8, Can I exploit your PureExo bundle (Cat. #P100) on our D.discoideum amoeba?
A: PureExo exosome isolation bundle captures launched exosomes or microvesicles from cells. For amoeba exosome isolation, we suggest to assemble amoeba custom medium because the an identical technique to assemble cell custom medium.

9, What is the composition of the buffer that the last word exosomes prep may be in? Will it have an effect on my downstream assay?
A: The final word isolated exosomes are aggregated nano-particles blended with a very small amount of genuine custom medium or serum. PureExo Exosome Isolation bundle provides intact exosomes which allows for a kinds of downstream functions along with exosome flooring labeling10， How have you ever learnt that an exosome prep is pure, free of completely different membrane-derived microparticles
(shredding vesicles)?
A: The isolated microvesicle from cell custom medium or serum using our isolation kits confirmed sphere membrane encapsulated particles with the diameters varied between 20-200nm beneath EM scanning. These characters help to seek out out that the harvested microvesicle are exosomes. Shedding particles are reported to be irregular type with diameter as a lot as 1000nm. In addition to, the size distribution pattern is confirmed by dynamic mild scattering indicating the isolated microvesicles are of diameter between 100 and 250nm.

11， Are exosome functionally energetic?
A: Our collaborators used our kits to detect specific mirRNAs carried by exosomes. Luciferase train assay
confirmed energetic function of exosomal microRNA after the exosomes had been utilized to cells transfected with specializing in mRNA 3’UTR vector.

12， Is it very important to utilize glass tubes?
A: Using glass tubes instead of plastic tubes is required in step Three and step 4.
Please uncover glass tubes from FisherScientific. http://www.fishersci.com (Look for Fisherbrand* Disposable
Borosilicate Glass Tubes Cat. NO: 14-961-26).

13， Why can’t I see clear fluff middle layer?
A: The subsequent coping with errors may end result within the failure:
1). Decision A/B/C did not mix absolutely sooner than together with the cell custom medium or serum.
2). Did not perform a low velocity centrifugation to remove cells and cell particles.
3). The ratio is off. As an illustration: 0.125 mL A + 0.125 mL B + 0.5mL C to 2 mL medium. Overproportion of serum/medium amount to Decision mixture (A/B/C) will lead to an indistinctive interphase.
4). The utmost starting amount is 4 ml medium, and it have to be from half million cultured cells or a lot much less. If cell amount or medium amount exceed this amount, the sample have to be splits into various samples to proceed.
5). Cultured cells weren’t starve for 48 hours to remove contaminated exosomes derived from FBS.

14， What biomarker(s) do you utilize to find out / characterize the isolated exosomes?
A: We use exosome markers akin to flotillin, CD9, CD63, Alix, and TSG101 and lots of others to characterize the isolated exosomes.

15， What is the yield of exosome using your kits?
A: For P100 bundle (PureExo bundle for cell custom medium), normally, 50 ~ 100 ul exosomes will likely be isolated from 2ml cell custom media (As a lot as 4 ml of medium will likely be processed per response). If the entire harvested exosomes are used for exosomal protein extraction (using our P200 bundle), your complete protein yield ranges from 150 ~ 200 ug / 2mL medium, 300 ~ 400 ug / 4 ml medium. If the entire isolated exosome are used for RNA extraction (using our P200 bundle), entire exosomal RNA yield is 50 ~ 100 ng / 2ml medium, 100 ~ 200 ng / 4ml medium. For P101 bundle (PureExo bundle for serum or plasma), normally 100 ~ 200 ul exosome will likely be isolated from 100ul to 600ul serum. If the entire harvested exosomes are used for exosomal protein extraction (using our P200 bundle), your complete exosomal protein yield ranges from 300-400 ug protein. If the entire isolated exosome are used for RNA extraction (using our P200 bundle), entire exosomal RNA yield is 100 ~ 300ng exosomal RNA For P120 bundle (DiagExo bundle for Urine), normally 50 ~ 200 ul exosome will likely be isolated from Three mL urine. If the entire harvested exosomes are used for exosomal protein extraction (using our P200 bundle), your complete exosomal protein yield ranges from 150-400ug protein. If the entire isolated exosome are used for RNA extraction (using our P200 bundle), entire exosomal RNA yield is 50 ~ 200ng exosomal RNA

16, Can I exploit heparin or EDTA tube to assemble blood sample if I need to isolate exosome from plasma?
A: No. Heparin and EDTA will significantly impair the downstream assays. Use No-heparin-No-EDTA tube to assemble the blood sample. Centrifuge the sample immediately to assemble the plasma for exosome isolation. .