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Gentaur - Antibodies, ELISA kits, PCR, Cell culture, Kronos Dio, Bullet Blender

Exo-FBS™ Exosome-depleted FBS

Exosomes are 60-120 nm membrane vesicles secreted by most cell types in vivo and in vitro. Exosomes are found in blood, urine, amniotic fluid, malignant ascite fluids, urine and in media from cells in culture. Exosomes contain distinct subsets of microRNAs depending upon the cell type from which they are secreted. Standard growth medium for most cells in culture require fetal bovine serum (FBS) as a growth supplement to DMEM. FBS is derived from bovine (cow) serum and contains a high abundance of cow exosome vesicles. These exosomes can interfere or cause significant background issues when studying the exosomes secreted from your cells of interest in standard culture conditions.

SBI has developed an exosome-depleted FBS growth supplement called Exo-FBS that has been stripped of bovine exosomes. Exo-FBS supports equivalent growth of many types of cells in culture, is devoid of cow CD63 positive exosomes and does not have any measurable bovine microRNAs. Perform your studies of cellular secreted exosomes in culture without the worry of contaminating cow exosomes in your experiments. No ultracentrifugation required. Available as standard FBS supplement or heat inactivated FBS media supplement (treated at 65°C for 15 minutes before bovine exosome removal).

  • Exosome-sized vesicles removed
  • FBS that has been stripped of CD63 positive cow exosomes
  • No detectable cow microRNAs
  • Same cellular growth rates supported as standard FBS
  • Use the same as standard FBS (add 10% in DMEM or RPMI)

 

NanoSight particle analysis of FBS and Exo-FBS

Standard FBS and Exo-FBS samples were diluted 1:1000 and then analyzed for particle size and abundance using a NanoSight LM10 instrument. The standard FBS sample shows a significant amount of exosome-sized microvesicles where the Exo-FBS exosome-depleted FBS sample has a drastic reduction in exosome particles.

 

 

Exo-FBS is depleted of bovine CD63 exosomes

The tetraspanin CD63 protein is a common marker for exosomes. We utilized a bovine-specific anti-CD63 antibody to develop an Enzyme Linked Immunosorbent Assay (ELISA). Equal volumes (50 ul) of either standard FBS or Exo-FBS depleted media supplement were used in this ELISA assay. Amounts of CD63-positive bovine exosomes dramatically reduced. The results are graphed and normalized to the signal level of standard FBS.

 

 

 

 

 

Bovine microRNAs are absent in Exo-FBS

Standard FBS and Exo-FBS media supplements (4 ml) were treated with Trizol extraction methods to recover exosome RNAs. RNA was converted to cDNA and 72 individual bovine microRNAs were measured by qPCR using SBI's QuantiMir system. Of the 72 microRNAs tested, 12 yielded amplification curves in the FBS sample but not in the Exo-FBS sample.

Bovine microRNAs present in FBS are no longer detectable in Exo-FBS - No more cow microRNAs!

Exo-FBS media supports robust growth of cells in culture equal to standard FBS media

Complete medium either with 10% standard FBS or with 10% Exo-FBS supplement. HT1080 fibrosarcoma cells, PC-3 prostate cancer cells, MCF-7 breast cancer cells and HEK293 cells were seeded at either 10,000 or 20,000 cells and then cultured under standard conditions at 37°C with 5% CO2 for 5 days in the medium indicated. The cells were imaged for growth numbers and morphologies. Equivalent growth and morphologies were observed for standard FBS and Exo-FBS media tested across these 4 cell lines tested.



Discover microRNA and Protein Biomarkers in Patient Biofluids
Exosomes are 60 - 120 nm membrane vesicles secreted by most cell types and contain distinct subsets of microRNAs depending upon the tumor from which they are secreted. The ExoQuick exosome precipitation reagent makes microRNA and protein biomarker discoveries simple, reliable and quantitative.

ExoQuick benefits
• No time-consuming ultracentrifugation
• Less expensive than costly Antibodies and beads
• No complicated syringes required
• Compatible with biofluid from any species
• More effective than any other method
• Isolate intact exosomes for functional studies
• Use as little as 100 µl of serum
• Start with 5 or 10 ml Media or Urine and use ExoQuick-TC

Precipitate Exosomes from Serum with ExoQuick
EXOQ5A-1 Exosome precipitation solution 5ml 409 EUR
EXOQ20A-1 Exosome precipitation solution 20ml 1210 EUR

Isolate Exosomes from Culture Media and Urine with ExoQuick-TC
EXOTC10A-1 ExoQuick TC 10 ml 408 EUR
EXOTC50A-1 ExoQuick TC 50 ml 1253 EUR




Exosome Graphics ExoTC_1

Discover microRNA and Protein Biomarkers in Patient Biofluids
Exosomes are 40 –100 nm membrane vesicles secreted by most cell types in vivo and in vitro. Exosomes are found in blood, urine, amniotic fluid, malignant ascite fluids and contain distinct subsets of microRNAs depending upon the tumor from which they are secreted. SBI's ExoQuick exosome precipitation reagent makes microRNA and protein biomarker discoveries simple, reliable and quantitative. Enrich for circulating exosomal microRNAs with ExoQuick and accurately profile them using SBI’s QuantiMir qPCR arrays.

* No time-consuming ultracentrifugation
* Less expensive than costly Antibodies and beads
* No complicated syringes required
* Compatible with biofluid from any species
* More effective than any other method
* Isolate intact exosomes for functional studies
* Use as little as 100 µl of serum or biofluid

Isolate Exosomes from Culture Media and Urine with ExoQuick-TC

Discover microRNA and Protein Biomarkers in Patient Biofluids
Exosomes are 40 - 100 nm membrane vesicles secreted by most cell types in vivo and in vitro. Exosomes are found in blood, urine, amniotic fluid, malignant ascite fluids and contain distinct subsets of microRNAs depending upon the tumor from which they are secreted. SBI's ExoQuick exosome precipitation reagent makes microRNA and protein biomarker discoveries simple, reliable and quantitative. Enrich for circulating exosomal microRNAs with ExoQuick and accurately profile them using SBI's SeraMir™ qPCR arrays.

ExoQuick benefits

• No time-consuming ultracentrifugation
• Less expensive than costly Antibodies and beads
• No complicated syringes required
• Compatible with biofluid from any species
• More effective than any other method
• Isolate intact exosomes for functional studies
• Use as little as 100 µl of serum
• Start with 5 or 10 ml Media or Urine and use ExoQuick-TC

How to use ExoQuick

How to use ExoQuick

Amounts of ExoQuick to add to various biofluids

Amounts of ExoQuick to add to various biofluids

ExoQuick exosomes can be transfered between cells

SBI created a stable 293TN cell line overexpressing the Cyto-Tracer™ pCT-CD63-GFP fusion protein (catalog# CYTO120-PA-1). The media from the cells was collected 48 h after plating and the exosomes from the media were precipitated using ExoQuick-TC. The exosome pellet recovered was resuspended in 30ul PBS and 10ul was added to newly plated HT1080 cells. HT1080 cells were visualized 72 h after the addition of the CD63-GFP labeled exosomes and then re-plated. Following another 24h, the cells were again visualized for GFP fluorescence and imaged. The exosomes appear to dock with the cells within 72 h and some are found to be internalized after 96 h.

ExoQuick exosomes can be transfered between cells

Analysis of ExoQuick Serum and Urine exosomes using the NanoSight LM10

The NanoSight LM10 instrument is based on a conventional optical microscope and uses a laser light source to illuminate nano-scale particles within a 0.3 ml sample introduced to the viewing unit with a disposable syringe. Enhanced by a near perfect black background, particles appear individually as point-scatterers moving under Brownian motion. The image analysis Nanoparticle Tracking Analysis (NTA) software suite allows users to automatically track and size nanoparticles on an individual basis. Results are displayed as a frequency size distribution graph and output to spreadsheet. In addition, video clips of images may be captured and archived for future reference.

ExoQuick serum exosome analysis

Normal human serum from 50 pooled samples was used. Only 250ul serum was combined with 63ul ExoQuick to pellet the exosomes in 30 minutes. The exosome pellet was resuspended in 100ul PBS, diluted 1:10,000 and visualized on the NanoSight LM10 instrument. The analysis shows that ExoQuick isolated 90nm exosomes with a recovery of 2.74 x 10^12 particles/ml.

ExoQuick-TC conditioned media exosome analysis

Human HT1080 lung sarcoma cell line was cultured in conditioned media (serum-free) for 72 hours. Ten milliliters of the media was combined with 2ml ExoQuick-TC to pellet the exosomes overnight. The exosome pellet was resuspended in 1ml PBS, diluted 1:40 and visualized on the NanoSight LM10 instrument. The analysis shows that ExoQuick isolated 133nm exosomes with a recovery of 1.74 x 10^9 particles/ml.

 

Product Portfolio

  • Lentiviral technologies
  • Molecular Tools
  • Stem cell technologies
  • PiggyBac Transposon
  • Minicircle Technology
  • MicroRNA research tools
  • RNAi Libraries
  • Molecular tools
  • Gene Analysis

 

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