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ExoQuick-TC Fast, scalable exosome isolation from tissue culture media
货号：EXOTC10A-1

ExoQuick-TC’s fast, ultracentrifugation-free method:

• Saves time and labor
• Is easily scalable
• Conserves precious sample
• Delivers high yields of functional, high quality exosomes
• Can be used to isolate exosomes for a wide range of downstream applications, including
  • Biomarker studies
  • Exosomal miRNA profiling
  • Exosomal proteomics
  • Exosomal lipidomics/metabolomics
  • Functional studies, such as in cell-to-cell signaling
  • Basic biology, such as role in tumorigenesis

ExoQuick-TC is a proprietary polymer that gently precipitates exosomes. First, pre-clear your samples of cells and cellular debris, and then simply add the appropriate amount of ExoQuick-TC to your cleared biofluid, refrigerate, and centrifuge (see the product manual for protocol details). Your exosomes will be in the pellet, ready for resuspension in an appropriate solution.

Biofluid	Sample volume	ExoQuick-TC Volume
Tissue culture media, urine, cerebrospinal fluid (CSF), etc.	5 mL	1 mL

 

In electron microscopy studies, exosomes isolated with ExoQuick-TC appear similar to exosomes isolated using ultracentrifugation^1-2, and these exosomes are also active in numerous functional assays^1-3.

Exosomes isolated with ExoQuick-TC can be used for all types of protein profiling and protein characterization studies, such as mass spectrometry, Western blotting, ELISA, and more. Higher protein yields are achieved by ExoQuick purification than by chromatography, DynaBeads, or ultracentrifugation.

Exosomes isolated with ExoQuick-TC also provide excellent samples for studying exosome-associated nucleic acids such as microRNAs, siRNAs, and even mRNA. Quantitative analytical techniques such as qPCR, microarray studies, and next-generation sequencing are all compatible with nucleic acids isolated from ExoQuick-TC-purified exosomes.

Backed by a growing number of publications, ExoQuick-TC is often the best option for researchers working with low sample volumes, such as clinical research samples or small animal models.

ExoQuick-TC exosome isolation methods are patented technologies⁴.

Choose the right ExoQuick for your biofluid:

Catalog #	Product	Biofluid
EXOQ5A-1, EXOQ20A-1	The Original ExoQuick	For serum, plasma, and ascites fluid
EXOTC10A-1, EXOTC50A-1	ExoQuick-TC®	For all other biofluids
EXOLP5A-1	ExoQuick-LP	For removing contaminating lipoprotein particles from plasma or serum before ExoQuick precipitation
EXOQ5TM-1	ExoQuick Plasma Prep with Thrombin	For de-fibrinating plasma before exosome isolation for efficient recovery and high yields
EXOCG50A-1	ExoQuick-CG	For preparing exosomes used in pre-clinical in vivo applications
EQPL10A-1, EQPL10TC	ExoQuick PLUS and ExoQuick-TC PLUS	For sensitive applications such as mass spectrometry, exosome labeling, and in vivo/ex vivo exosome delivery

 

REFERENCES

1. Chugh PE, et al. Systemically Circulating Viral and Tumor-Derived MicroRNAs in KSHV-Associated Malignancies. PLoS Pathog. 2013. 9(7): e1003484. PMCID: PMC3715412.

1. Umezu T, et al. Leukemia cell to endothelial cell communication via exosomal miRNAs. Oncogene. 2013 May 30. 32(22):2747-55. PMID: 22797057.

1. Sohel MM, et al. Exosomal and Non-Exosomal Transport of Extra-Cellular microRNAs in Follicular Fluid: Implications for Bovine Oocyte Developmental Competence. PLoS One. 2013 Nov 4. 8(11):e78505. PMCID: PMC3817212.

1. Antes T, et al. Methods for Microvesicle Isolation and Selective Removal. Patent No.: US 9,005,888 B2.

 

How It Works

High-throughput, quantitative exosome recovery

ExoQuick-TC can be used to purify exosomes from a wide variety of tissue culture media⁴, and from certain biofluids such as saliva¹, urine², follicular fluid³, and breast milk⁵. With a simple workflow involving minimal hands-on time and low input sample volume requirements, ExoQuick-TC is an excellent option for researchers who need to purify multiple exosome samples.

To isolate exosomes from tissue culture media, simply:

• Add an appropriate volume of ExoQuick-TC
• Incubate overnight at 4°C
• Isolate exosomes with a 30-minute low-speed spin (1500g).

Isolated exosomes can be found in the pellet and resuspended in an appropriate solution.

You can verify the presence of exosomes with a number of different methods, including Western blotting for general exosome markers (CD63, CD9, CD81, and HSP70), NanoSight analysis, or EM (learn about different ways to detect exosomes and more in our Exosome Basics Guide).

The Bottom Line
With ExoQuick-TC, you can obtain high-quality exosomes from tissue culture media and certain biofluids using a protocol that can easily be performed on multiple samples and requires very low volumes of input sample.

REFERENCES

1. Yang J, et al. Detection of Tumor Cell-Specific mRNA and Protein in Exosome-Like Microvesicles from Blood and Saliva. PLoS ONE. 2014. 9(11): e110641. PMCID: PMC 4232306.

1. Alvarez ML. Isolation of urinary exosomes for RNA biomarker discovery using a simple, fast, and highly scalable method. Methods Mol Biol. 2014. 1182:145-70. PMID: 25055908.

1. Sohel MM, et al. Exosomal and Non-Exosomal Transport of Extra-Cellular microRNAs in Follicular Fluid: Implications for Bovine Oocyte Developmental Competence. PLoS One. 2013 Nov 4. 8(11):e78505. PMCID: PMC3817212.

1. Zhu L, et al. Novel method for extracting exosomes of hepatocellular carcinoma cells. World J Gastroenterol. 2014 June 7. 20(21): 6651-6657. PMCID: PMC4047354.

1. Gu Y, et al. Lactation-Related MicroRNA Expression Profiles of Porcine Breast Milk Exosomes. PLoS ONE. 2012. 7(8): e43691. PMCID: PMC3427246.

 

Supporting Data

Characterizing ExoQuick-TC exosomes with NanoSight

 

Exosomes purified with ExoQuick-TC from tissue culture media show the expected particle size distribution and high concentration yields when analyzed using NanoSight’s Nanoparticle Tracking Analysis (NTA, Figure 1).

Figure 3.Exosome size distribution and yields from tissue culture media. Cells from a human HT1080 lung sarcoma cell line were cultured in conditioned media (serum-free) for 72 hours. 10 mL of media was combined with 2 mL ExoQuick-TC, incubated overnight at 4°C, and centrifuged at 1500g for 30 minutes to isolate exosomes. The exosome pellet was resuspended in 1 mL PBS, diluted 1:40, and visualized on the NanoSight LM10 instrument. The analysis shows that ExoQuick-TC recovered 133 nm exosomes at a concentration of 1.74 x 10⁹ particles/mL.

 

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Product Documentation

• User Manual: ExoQuick-TC

Citations

• Jiang, XC & Gao, JQ. (2017) Exosomes as novel bio-carriers for gene and drug delivery. Int J Pharm. 2017 Apr 15; 521(1-2):167-175. PM ID: 28216464
• Selmaj, I, et al. (2017) Global Exosome Transcriptome Profiling Reveals Biomarkers for Multiple Sclerosis. Ann. Neurol.. 2017 Apr 15;. PM ID: 28411393
• Markowska, A, et al. (2017) A novel method for the isolation of extracellular vesicles and RNA from urine. Journal of Circulating Biomarkers. 2017 Jun 13;. Link: Journal of Circulating Biomarkers
• Lee, YS, et al. (2017) Exosomes derived from palmitic acid-treated hepatocytes induce fibrotic activation of hepatic stellate cells. Sci Rep. 2017 Jun 16; 7(1):3710. PM ID: 28623272
• Yang, Y, et al. (2017) Exosomes from Plasmodium-infected hosts inhibit tumor angiogenesis in a murine Lewis lung cancer model. Oncogenesis. 2017 Jun 26; 6(6):e351. PM ID: 28650446
• Caponnetto, F. (2017) Size-dependent internalization of exomemes. Thesis. ;. Link: Thesis
• Belo, R, et al. (2017) Leishmania infantum Exoproducts Inhibit Human Invariant NKT Cell Expansion and Activation. Front Immunol. 2017 Jul 4; 8:710. PM ID: 28674535
• Hubal, MJ, et al. (2017) Circulating adipocyte-derived exosomal MicroRNAs associated with decreased insulin resistance after gastric bypass. Obesity (Silver Spring). 2017 Jan 1; 25(1):102-110. PM ID: 27883272
• McNicholas, K & Michael, MZ. (2017) Immuno-characterization of Exosomes Using Nanoparticle Tracking Analysis. Methods Mol. Biol.. 2017 Dec 12; 1545:35-42. PM ID: 27943205
• Yang, Y, et al. (2017) MSCs-Derived Exosomes and Neuroinflammation, Neurogenesis and Therapy of Traumatic Brain Injury. Front Cell Neurosci. 2017 Mar 15; 11:55. PM ID: 28293177
• Broner, EC, et al. (2017) TSAP6 is a novel candidate marker of poor survival in metastatic high-grade serous carcinoma. Hum. Pathol.. 2017 Feb 1; 60:180-187. PM ID: 27825812
• Nakata, R, et al. (2017) Contribution of neuroblastoma-derived exosomes to the production of pro-tumorigenic signals by bone marrow mesenchymal stromal cells. Journal of Extracellular Vesicles. 2017 Jun 25; 6(1). Link: Journal of Extracellular Vesicles
• Ichinohe, N, et al. (2017) Transplantation of Thy1(+) Cells Accelerates Liver Regeneration by Enhancing the Growth of Small Hepatocyte-Like Progenitor Cells via IL17RB Signaling. Stem Cells. 2017 Apr 1; 35(4):920-931. PM ID: 27925343
• Cheng, L, et al. (2017) Exosomes from Melatonin Treated Hepatocellularcarcinoma Cells Alter the Immunosupression Status through STAT3 Pathway in Macrophages. Int. J. Biol. Sci.. 2017 Jun 28; 13(6):723-734. PM ID: 28655998
• Ocaña, GJ, et al. (2017) Inflammatory stress of pancreatic beta cells drives release of extracellular heat-shock protein 90α. Immunology. 2017 Jun 1; 151(2):198-210. PM ID: 28190264
• András, IE, et al. (2017) Extracellular vesicles of the blood-brain barrier: Role in the HIV-1 associated amyloid beta pathology. Mol. Cell. Neurosci.. 2017 Mar 1; 79:12-22. PM ID: 28040512
• Gong, M, et al. (2017) Mesenchymal stem cells release exosomes that transfer miRNAs to endothelial cells and promote angiogenesis. Oncotarget. 2017 Apr 1;. PM ID: 28423355
• Richards, KE, et al. (2017) Cancer-associated fibroblast exosomes regulate survival and proliferation of pancreatic cancer cells. Oncogene. 2017 Mar 30; 36(13):1770-1778. PM ID: 27669441
• Kamalden, TA, et al. (2017) Exosomal MicroRNA-15a Transfer from the Pancreas Augments Diabetic Complications by Inducing Oxidative Stress. Antioxid. Redox Signal.. 2017 Mar 17;. PM ID: 28173719
• Kastelowitz, N, et al. (2017) Peptides derived from MARCKS block coagulation complex assembly on phosphatidylserine. Sci Rep. 2017 Jun 27; 7(1):4275. PM ID: 28655899