Cellexus Limited Cell Culture Biosystems

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 probably be launched into the circulation system. So, if the exosomes from these kinds of samples are isolated, it’d incorporates Albumin.

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

9, What is the composition of the buffer that the last word exosomes prep could be in? Will it have an effect on my downstream assay?
A: The last word isolated exosomes are aggregated nano-particles blended with a extremely small amount of genuine custom medium or serum. PureExo Exosome Isolation package deal provides intact exosomes which allows for a varieties of downstream functions along with exosome ground labeling10， How have you ever learnt that an exosome prep is pure, free of totally 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 search 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 as a substitute of plastic tubes is required in step Three and step 4.
Please uncover glass tubes from FisherScientific. http://www.fishersci.com (Search for Fisherbrand* Disposable
Borosilicate Glass Tubes Cat. NO: 14-961-26).

13， Why can’t I see clear fluff heart layer?
A: The following coping with errors may outcome within the failure:
1). Decision A/B/C did not mix totally 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. For example: 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 end in 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 quite a few 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 use to find out / characterize the isolated exosomes?
A: We use exosome markers akin to flotillin, CD9, CD63, Alix, and TSG101 and plenty of others to characterize the isolated exosomes.

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