For the isolation of exosomes from human plasma, 250 µL of plasma was thawed on ice and prepared by removing platelets and large vesicles by spinning at 1,500 g and 10,000 g for 10 and 20 minutes, respectively. Two-hundred microlitres of prepared plasma was then mixed with 100 µL of precipitation reagent and incubated at 4°C for 5 minutes before pelleting exosomes at 20,000 g for 30 minutes. The supernatant was removed, and the exosome-containing pellet resuspended in 100 µL of PBS and purified on columns as before. The 200 µL exosome preparation was further filtered through an Ultrafree® 0.22 µm centrifugal filter device to remove any large contaminating vesicles.
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Exosomes
Exosomes
Exosomes are small extracellular vesicles released by cells that play a crucial role in intercellular communication.
These nanometer-sized membrane-enclosed particles carry a cargo of proteins, lipids, and genetic material that can be transferred to recipient cells, influencing their function and behavior.
Exosomes are involved in a variety of physiological and pathological processes, including immune response, tumor progression, and neurodegenerative disorders.
Reserach into exosomes is a rapidly growing field, offering new insights into cell-to-cell signaling and potential therapeutic applications.
PubCompare.ai's AI-driven platform can optimize your exosomes reserach by helping you locate the best protocols from literature, preprints, and patents, while utilizing AI-driven comparisons to enhance reproducibilty and accuracy.
Streamline your exosomes reserach with PubCompare.ai's powerful capabilities.
These nanometer-sized membrane-enclosed particles carry a cargo of proteins, lipids, and genetic material that can be transferred to recipient cells, influencing their function and behavior.
Exosomes are involved in a variety of physiological and pathological processes, including immune response, tumor progression, and neurodegenerative disorders.
Reserach into exosomes is a rapidly growing field, offering new insights into cell-to-cell signaling and potential therapeutic applications.
PubCompare.ai's AI-driven platform can optimize your exosomes reserach by helping you locate the best protocols from literature, preprints, and patents, while utilizing AI-driven comparisons to enhance reproducibilty and accuracy.
Streamline your exosomes reserach with PubCompare.ai's powerful capabilities.
Most cited protocols related to «Exosomes»
Blood Platelets
Exosomes
Homo sapiens
isolation
Medical Devices
Plasma
Strains
CCM was harvested from SK-MES-1 cells and centrifuged using a Beckman Coulter Allegra® X-15R centrifuge at 300 g at 4°C for 10 minutes to remove detached cells. Supernatant was collected and filtered through 0.22 µm filters (Merck Millipore) to remove contaminating apoptotic bodies, microvesicles and cell debris. Clarified CCM was then centrifuged in a Beckman Coulter Optima™ L-80XP Ultracentrifuge at 100,000 gavg at 4°C for 90 minutes with a Type 50.2 Ti rotor (k-factor: 157.7) to pellet exosomes. The supernatant was carefully removed, and crude exosome-containing pellets were resuspended in 1 mL of ice-cold PBS and pooled. A second round of ultracentrifugation [100,000 gavg at 4°C for 90 minutes with a Type 50.2 Ti rotor (k-factor: 157.7)] was carried out, and the resulting exosome pellet resuspended in 500 µL of PBS (Supplementary Fig. 1).
A 300
Allegra
Apoptotic Bodies
Cell-Derived Microparticles
Cells
Common Cold
Exosomes
Pellets, Drug
Ultracentrifugation
Fresh or previously frozen murine hemi-brains were dissected and treated with 20 units/ml papain (Worthington) in Hibernate E solution (3 ml/hemi-brain; BrainBits, Springfield, IL) for 15 min at 37 °C. The brain tissue was gently homogenized in 2 volumes (6 ml/hemi-brain) of cold Hibernate E solution. The brain homogenate was sequentially filtered through a 40-μm mesh filter (BD Biosciences) and a 0.2-μm syringe filter (Thermo Scientific). Exosomes were isolated from the filtrate as described previously (15 ). Briefly, the filtrate was sequentially centrifuged at 300 × g for 10 min at 4 °C, 2000 × g for 10 min at 4 °C, and 10,000 × g for 30 min at 4 °C to discard cells, membranes, and debris. The supernatant was centrifuged at 100,000 × g for 70 min at 4 °C to pellet exosomes. The exosome pellet was resuspended in 60 ml of cold PBS (Invitrogen), and the exosome solution was centrifuged at 100,000 × g for 70 min at 4 °C. The washed exosome pellet was resuspended in 2 ml of 0.95 m sucrose solution and inserted inside a sucrose step gradient column (six 2-ml steps starting from 2.0 m sucrose up to 0.25 m sucrose in 0.35 m increments, with the 0.95 m sucrose step containing the exosomes). The sucrose step gradient was centrifuged at 200,000 × g for 16 h at 4 °C. One-ml fractions were collected from the top of the gradient, and fractions flanking the interphase separating two neighboring sucrose layers were pooled together for a total of seven fractions (a, top 1-ml fraction; b, 2-ml; c, 2-ml; d, 2-ml; e, 2-ml; f, 2-ml; and g, bottom 1-ml fraction). These fractions were diluted in cold PBS and centrifuged at 100,000 × g at 4 °C for 70 min. Sucrose gradient fraction pellets were resuspended in 20 μl of cold PBS. Two μl were used to measure acetylcholine esterase (AChE) activity, and 2-μl were used for EM. Exosome lysate was prepared by mixing 16 μl of the leftover solution with an equal volume of 2× radioimmune precipitation assay lysis buffer supplemented with a mixture of protease inhibitors. We used 2 μl of the lysate to quantify exosomal protein content (BCA protein assay kit, Pierce) and 10 μl of the lysate (31% of the exosome lysate total volume) for protein analysis by Western blotting.
Acetylcholinesterase
Biological Assay
Brain
Buffers
Cells
Cold Temperature
Exosomes
Freezing
Interphase
Mus
Papain
Pellets, Drug
Protease Inhibitors
Proteins
Sucrose
Syringes
Tissue, Membrane
Tissues
Western Blot
Frozen plasma specimens were thawed and centrifuged at 2,000×g for 10 min at 4°C and then at 10,000–14,000×g for 30 min at 4°C. Clarified plasma was passed through 0.22 µm-pore Millipore filter and used for exosome isolation by SEC performed using 1.5 cm×12 cm mini-columns (Bio-Rad, Hercules, CA, USA; Econo-Pac columns) packed with Sepharose 2B (Sigma-Aldrich, St. Louis, MO, USA). The column bed volume is 10 mL. Prior to applying clarified plasma, the column is washed with 20 mL of phosphate-buffered saline (PBS), and a porous frit is placed at the top of the gel to prevent its disturbance during subsequent elution with PBS. Clarified plasma (0.5–1.0 mL) was loaded onto the column and five 1 mL fractions corresponding to the void volume peak were collected. Fractions # 3, #4 and #5 were tested for protein content, morphology by transmission electron microscopy (TEM) and in functional assays. In preparation for western blots, the fractions were concentrated using 300,000 MWCO VivaSpin 500 Centrifugal Concentrators (Sartorius Corp, New York, NY, USA) by centrifugation at 5,000×g for 2–15 min, depending on the content. Supplementary Fig. 1 shows the schema for isolation of plasma exosomes by the mini-SEC method.
Biological Assay
Centrifugation
Exosomes
Freezing
isolation
Phosphates
Plasma
Proteins
Saline Solution
Sepharose
Transmission Electron Microscopy
Urination
Western Blot
We manually curated RNA-seq data for samples derived from human blood exosomes from the NCBI Gene Expression Omnibus (GEO, http://www.ncbi.nlm.nih.gov/geo/ ) (14 (link)) or Short Read Archive (SRA, http://www.ncbi.nlm.nih.gov/sra/ ) (15 (link)). Currently, exoRBase includes 87 exosome samples from six experiments (GSE100206, GSE99985, GSE100063, GSE100207, GSE100232, GSE93078). These samples are from different biological conditions, including normal persons (NP), coronary heart disease (CHD), colorectal cancer (CRC), hepatocellular carcinoma (HCC), pancreatic adenocarcinoma (PAAD) and Breast Cancer (BC). In addition, the current collections include whole blood experiments (five samples, GSE73570). To complement the exoRBase database, we also searched PubMed for exosomal RNAs with experimental validation, which resulted in 18 literature records involving 77 exosomal RNAs. The RNA-seq expression profiles of 30 tissues from the GTEx project (13 (link)) were used to annotate possible original tissues of exosomal RNAs. In addition, resource of all human circRNAs were downloaded from circBase database (16 (link)). The genomic coordinates of circRNAs were converted to hg38 using the UCSC liftover tool. Then these circRNAs were combined with our collections. We also integrated the tissue types of expressed circRNAs from the results of our previous research, where 27 296 circRNAs were detected from six tissues (11 (link)).
Adenocarcinoma
Biopharmaceuticals
BLOOD
Colorectal Carcinoma
Exosomes
Gene Expression
Genome
Heart Disease, Coronary
Hepatocellular Carcinomas
Histocompatibility Testing
Homo sapiens
Malignant Neoplasm of Breast
Manpower
Pancreas
RNA
RNA, Circular
RNA-Seq
Tissues
Most recents protocols related to «Exosomes»
Example 12
The binding of CIBN and CRY2 in cells expressing CIBN-EGFP-CD9 and JNK-mCherry-Cry2 at 488 nm wavelength blue light, and the loading of JNK within the exosome is evaluated.
For the massive production of JNK-loaded exosomes, cells stably expressing CIBN-EGFP-CD9 gene and JNK-mCherry-CRY2 gene are established, and exosomes are isolated and purified by Tangential Flow Filtration (TFF) method from culture supernatant.
Functional analysis of JNK-loaded exosomes is performed in target cells:
Target cells are treated with the JNK-loaded exosomes to show the functional activity.
Animal models are administered with the JNK-loaded exosomes by i.p. or i.v. to show therapeutic effect.
Animal Model
Cells
Exosomes
Filtration
Genes
Kinase, Janus
Light
Therapeutic Effect
Example 7
Since the genetic material may be delivered by the exosomes, it has been hypothesized that the exosomes secreted from the cells cultured in a human ES medium may change the properties of surrounding cells or untreated cells. To verify this, the exosomes were extracted from the 2-day cultured medium of cells treated with ultrasonic wave cultured in the human ES medium environment, and the exosome extract was mixed and cultured for 6 days in a process of culturing the untreated cells in the human ES medium and a fibroblast culture medium, DMEM.
As a result, spheroid was produced in a group added with exosomes (
Cell Culture Techniques
Culture Media
Exosomes
Fibroblasts
Genetic Materials
Homo sapiens
Human Embryonic Stem Cells
Obstetric Delivery
Physical Stimulation
POU5F1 protein, human
Ultrasonic Waves
Example 11
The present inventors confirmed the binding of CIBN and CRY2 in cells expressing CIBN-EGFP-CD9 and PTEN-Cry2 at 488 nm wavelength blue light, and the loading of PTEN within exosome.
For the massive production of PTEN-loaded exosomes, cells stably expressing CIBN-EGFP-CD9 gene and PTEN-CRY2 gene were established (
Functional analysis of PTEN kinase-loaded exosomes is performed in target cells
Treatment of PTEN-loaded exosomes to target cells shows the functional activity.
Administration of PTEN-loaded exosomes by i.p. or i.v. to animal model shows therapeutic effect.
Animal Model
Cells
Exosomes
Filtration
Genes
Inventors
Light
Phosphotransferases
PTEN Phosphohydrolase
PTEN protein, human
Therapeutic Effect
Example 18
The binding of CIBN and CRY2 in cells expressing CIBN-EGFP-CD9 and Tbx18-mCherry-Cry2 at 488 nm wavelength blue light, and the loading of Tbx18 within the exosome is evaluated.
For the massive production of Tbx18-loaded exosomes, cells stably expressing CIBN-EGFP-CD9 gene and Tbx18-mCherry-CRY2 gene are established, and exosomes are isolated and purified by Tangential Flow Filtration (TFF) method from culture supernatant.
Functional analysis of Tbx18-loaded exosomes is performed in target cells:
Target cells are treated with the Tbx18-loaded exosomes to show the functional activity.
Animal models are administered with the Tbx18-loaded exosomes by i.p. or i.v. to show therapeutic effect.
Animal Model
Cells
Exosomes
Filtration
Genes
Light
Therapeutic Effect
Transcription Factor
Example 17
The present inventors confirmed the binding of CIBN and CRY2 in cells expressing CIBN-EGFP-CD9 and MyoD-mcherry-Cry2 at 488 nm wavelength blue light (
For the massive production of MyoD-loaded exosomes, cells stably expressing CIBN-EGFP-CD9 gene and MyoD-mcherry-CRY2 gene were established, and exosomes were isolated and purified by Tangential Flow Filtration (TFF) method from culture supernatant.
Treatment of MyoD-loaded exosomes to target cells showed the functional activity (
Cells
Exosomes
Filtration
Genes
Inventors
Light
Top products related to «Exosomes»
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TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
Sourced in United States, Lithuania, Japan
The Total Exosome Isolation Reagent is a laboratory product designed to isolate extracellular vesicles, including exosomes, from various biological samples such as cell culture media, blood, or other bodily fluids. The reagent utilizes a precipitation-based method to capture and concentrate these vesicles, facilitating their further analysis or downstream applications.
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The NanoSight NS300 is a nanoparticle characterization instrument. It uses Nanoparticle Tracking Analysis (NTA) technology to measure the size and concentration of particles in liquid suspensions. The instrument can analyze particles in the size range of 10 to 2,000 nanometers.
Sourced in United States, China, Denmark, Canada
ExoQuick Exosome Precipitation Solution is a reagent designed to precipitate and concentrate exosomes from biological samples, such as cell culture media or body fluids. The solution enables the rapid isolation of exosomes without the need for specialized equipment or lengthy ultracentrifugation procedures.
Sourced in United States, Canada, Lithuania
The Total Exosome Isolation Kit is a product designed to effectively isolate exosomes from various biological samples, such as cell culture media or biological fluids. The kit utilizes a proprietary precipitation reagent to extract exosomes, which can then be further analyzed or characterized using downstream applications.
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The MiRNeasy Mini Kit is a laboratory instrument designed for the extraction and purification of microRNA (miRNA) and other small RNA molecules from a variety of sample types, including cells, tissues, and body fluids. The kit utilizes a silica-membrane-based technology to facilitate the efficient isolation of high-quality miRNA and small RNA.
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The Agilent 2100 Bioanalyzer is a lab instrument that provides automated analysis of DNA, RNA, and protein samples. It uses microfluidic technology to separate and detect these biomolecules with high sensitivity and resolution.
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PKH26 is a fluorescent cell linker that binds to the lipid bilayers of cell membranes. It is used for labeling and tracking cells in various biological applications.
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The BCA Protein Assay Kit is a colorimetric detection and quantification method for total protein concentration. It utilizes bicinchoninic acid (BCA) for the colorimetric detection and quantification of total protein. The assay is based on the reduction of Cu2+ to Cu1+ by protein in an alkaline medium, with the chelation of BCA with the Cu1+ ion resulting in a purple-colored reaction product that exhibits a strong absorbance at 562 nm, which is proportional to the amount of protein present in the sample.
More about "Exosomes"
Exosomes are small, membrane-bound extracellular vesicles (EVs) that are released by cells and play a crucial role in intercellular communication.
These nanometer-sized particles, also known as 'exosome-like vesicles' or 'microvesicles', carry a cargo of proteins, lipids, and genetic material (including mRNA and miRNA) that can be transferred to recipient cells, influencing their function and behavior.
Exosomes are involved in a variety of physiological and pathological processes, such as immune response, tumor progression, and neurodegenerative disorders.
Research into exosomes is a rapidly growing field, offering new insights into cell-to-cell signaling and potential therapeutic applications.
To isolate and characterize exosomes, researchers often use techniques like TRIzol reagent for RNA extraction, FBS (fetal bovine serum) as a source of exosomes, and Total Exosome Isolation Reagent or ExoQuick Exosome Precipitation Solution for purification.
Nanoparticle tracking analysis using the NanoSight NS300 system can be used to measure the size and concentration of exosomes, while the Agilent 2100 Bioanalyzer provides insights into their molecular composition.
Exosome-associated proteins, such as those identified by the BCA protein assay kit, and the lipophilic dye PKH26 can be used to label and track exosomes.
The MiRNeasy Mini Kit is a common tool for extracting and purifying exosomal RNA for further analysis.
By leveraging PubCompare.ai's AI-driven platform, researchers can optimize their exosome studies by identifying the best protocols from literature, preprints, and patents, while using AI-driven comparisons to enhance reproducibility and accuracy.
This can help streamline the exosome research process and unlock new insights into this rapidly evolving field.
These nanometer-sized particles, also known as 'exosome-like vesicles' or 'microvesicles', carry a cargo of proteins, lipids, and genetic material (including mRNA and miRNA) that can be transferred to recipient cells, influencing their function and behavior.
Exosomes are involved in a variety of physiological and pathological processes, such as immune response, tumor progression, and neurodegenerative disorders.
Research into exosomes is a rapidly growing field, offering new insights into cell-to-cell signaling and potential therapeutic applications.
To isolate and characterize exosomes, researchers often use techniques like TRIzol reagent for RNA extraction, FBS (fetal bovine serum) as a source of exosomes, and Total Exosome Isolation Reagent or ExoQuick Exosome Precipitation Solution for purification.
Nanoparticle tracking analysis using the NanoSight NS300 system can be used to measure the size and concentration of exosomes, while the Agilent 2100 Bioanalyzer provides insights into their molecular composition.
Exosome-associated proteins, such as those identified by the BCA protein assay kit, and the lipophilic dye PKH26 can be used to label and track exosomes.
The MiRNeasy Mini Kit is a common tool for extracting and purifying exosomal RNA for further analysis.
By leveraging PubCompare.ai's AI-driven platform, researchers can optimize their exosome studies by identifying the best protocols from literature, preprints, and patents, while using AI-driven comparisons to enhance reproducibility and accuracy.
This can help streamline the exosome research process and unlock new insights into this rapidly evolving field.