Four Which package ought to I exploit for my urine and plasma samples?
A: Use DiagExo® package (Cat.#: P120) to isolate exosome from urine and Use PureExo® package for serum (Cat.#: P101) to isolate exosome from serum.

5, Can I exploit your package to isolate exosome or endothelial microparticles from cultured cells and from plasma?
A: Our P100 package can be utilized to isolate exosome from both cell tradition medium or serum. It’s not for cells.

6 Is your exosome prep contaminated with IgG and/or beads?
A: We use particular reagents to seize exosomes. We didn’t use antibody or beads for the isolation. So the ultimate exosome prep just isn’t contaminated with synthetic IgG or beads.

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

8, Can I exploit your PureExo package (Cat. #P100) on our D.discoideum amoeba?
A: PureExo exosome isolation package captures launched exosomes or microvesicles from cells. For amoeba exosome isolation, we recommend to gather amoeba tradition medium as the identical strategy to gather cell tradition medium.

9, What’s the composition of the buffer that the ultimate exosomes prep can be in? Will it affect my downstream assay?
A: The ultimate remoted exosomes are aggregated nano-particles blended with a really small quantity of authentic tradition medium or serum. PureExo Exosome Isolation package gives intact exosomes which permits for a types of downstream purposes together with exosome floor labeling10， How have you learnt that an exosome prep is pure, freed from different membrane-derived microparticles
(shredding vesicles)?
A: The remoted microvesicle from cell tradition medium or serum utilizing our isolation kits confirmed sphere membrane encapsulated particles with the diameters various between 20-200nm beneath EM scanning. These characters assist to find out that the harvested microvesicle are exosomes. Shedding particles are reported to be irregular form with diameter as much as 1000nm. As well as, the dimensions distribution sample is confirmed by dynamic gentle scattering indicating the remoted microvesicles are of diameter between 100 and 250nm.

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

12， Is it vital to make use of glass tubes?
A: Utilizing glass tubes as an alternative of plastic tubes is required in step Three and step 4.
Please discover glass tubes from FisherScientific. http://www.fishersci.com (Seek for Fisherbrand* Disposable
Borosilicate Glass Tubes Cat. NO: 14-961-26).

13， Why can’t I see clear fluff center layer?
A: The next dealing with errors could result in the failure:
1). Resolution A/B/C didn’t combine fully earlier than including the cell tradition medium or serum.
2). Didn’t carry out a low velocity centrifugation to take away cells and cell particles.
3). The ratio is off. For instance: 0.125 mL A + 0.125 mL B + 0.5mL C to 2 mL medium. Overproportion of serum/medium quantity to Resolution combination (A/B/C) will result in an indistinctive interphase.
4). The utmost beginning quantity is Four ml medium, and it must be from half million cultured cells or much less. If cell quantity or medium quantity exceed this quantity, the pattern must be splits into a number of samples to proceed.
5). Cultured cells weren’t starve for 48 hours to take away contaminated exosomes derived from FBS.

14， What biomarker(s) do you employ to determine / characterize the remoted exosomes?
A: We use exosome markers akin to flotillin, CD9, CD63, Alix, and TSG101 and many others to characterize the remoted exosomes.

15， What’s the yield of exosome utilizing your kits?
A: For P100 package (PureExo package for cell tradition medium), usually, 50 ~ 100 ul exosomes will be remoted from 2ml cell tradition media (As much as Four ml of medium will be processed per response). If all of the harvested exosomes are used for exosomal protein extraction (utilizing our P200 package), the entire protein yield ranges from 150 ~ 200 ug / 2mL medium, 300 ~ 400 ug / Four ml medium. If all of the remoted exosome are used for RNA extraction (utilizing our P200 package), whole exosomal RNA yield is 50 ~ 100 ng / 2ml medium, 100 ~ 200 ng / 4ml medium. For P101 package (PureExo package for serum or plasma), usually 100 ~ 200 ul exosome will be remoted from 100ul to 600ul serum. If all of the harvested exosomes are used for exosomal protein extraction (utilizing our P200 package), the entire exosomal protein yield ranges from 300-400 ug protein. If all of the remoted exosome are used for RNA extraction (utilizing our P200 package), whole exosomal RNA yield is 100 ~ 300ng exosomal RNA For P120 package (DiagExo package for Urine), usually 50 ~ 200 ul exosome will be remoted from Three mL urine. If all of the harvested exosomes are used for exosomal protein extraction (utilizing our P200 package), the entire exosomal protein yield ranges from 150-400ug protein. If all of the remoted exosome are used for RNA extraction (utilizing our P200 package), whole exosomal RNA yield is 50 ~ 200ng exosomal RNA

16, Can I exploit heparin or EDTA tube to gather blood pattern if I must isolate exosome from plasma?
A: No. Heparin and EDTA will considerably impair the downstream assays. Use No-heparin-No-EDTA tube to gather the blood pattern. Centrifuge the pattern instantly to gather the plasma for exosome isolation. .