Exosome Standards (HT29 cell line) Summary
| Description |
Highly pure, lyophilized exosome standards with superior stability, optimal for multiple applications including: Assay calibration, Spike-in control for exosome quantification, Protein marker analysis for different techniques such as Western Blot and Flow Cytometry, Extraction and analysis of exosomal RNA and DNA, Standardized positive controls for evaluating immunocapture performance.
Quantity per vial of 30 ug size (number of particles in 30 ug: > 1x10^8). Quantity per vial of 100 ug size (number of particles in 100 ug: > 1x10^10). Store lyophilized exosomes up to 3 years at 4C (Expire date indicated on the label). For reconstituted exosomes store at -20C for up to one month or at -80C for up to 6 months. Avoid repeated freeze-and-thraw cycles. |
Preparation
Method |
Isolation involves Tangential flow filtration combined with Size Exclusion Chromatogrpahy. Exosomes (small EVs) are quantified and validated for protein content and particle number by Nanoparticle Tracking Analysis as well as for common tetraspanins maker validation. Lyophilization does not alter stability of exosome proteins and nucleic acids. |
Applications/Dilutions
| Dilutions |
- Electron Microscopy
- ELISA
- Flow Cytometry
- Nucleic Acid Extraction
- Simple Western
|
Packaging, Storage & Formulations
| Storage |
Store at 4C. After reconstitution store at -70C. |
| Buffer |
Lyophilized from cell culture media |
| Concentration |
LYOPH |
| Reconstitution Instructions |
Add deionized water, 100 ul for Standard 100 ug and 30 ul for Standard 30 ug, to get a final concentration of 1 mg/mL. Resuspend exosomes pipetting the solution up and down 10-15 times, avoiding bubbles. Vortex the reconstituted standard for 60 seconds. |
Background
Exosomes are small endosome derived lipid nanoparticles (30-150nm) actively secreted by exocytosis by most living cells. Exosome release occurs either constitutively or upon induction, under both normal and pathological conditions, in a dynamic, regulated and functionally relevant manner. Both amount and molecular composition of released exosomes depend on the state of a parent cell. Exosomes have been isolated from diverse cell lines (hematopoietic cells, tumor lines, primary cultures, virus infected cells) as well as from biological fluids in particular blood (e.g. serum and plasma from cancer patients) and other body fluids (bronchoalveolar lavage fluid, pleural effusions, synovial fluid, urine, amniotic fluid, semen, saliva etc). Exosomes have pleiotropic physiological and pathological functions and an emerging role in diverse pathological conditions such as cancer, infectious and neurodegenerative diseases.
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Support products are
guaranteed for 6 months from date of receipt.
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Product General Protocols
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Blogs on Exosome Standards (HT29 cell line)
![Flow Cytometry: Exosome Standards (HT29 cell line) [NBP3-11685] - Phenotyping assays by FACS. Reconstituted Exosomes can be used for profiling biomarkers by FACS analysis. Recommended quantity: 5 ug of reconstituted Exosomes Standards for each test](http://images.novusbio.com/fullsize/Exosome-Standards-HT29-cell-line-Flow-Cytometry-NBP3-11685-img0001.jpg "Flow Cytometry: Exosome Standards (HT29 cell line) [NBP3-11685] - Phenotyping assays by FACS. Reconstituted Exosomes can be used for profiling biomarkers by FACS analysis. Recommended quantity: 5 ug of reconstituted Exosomes Standards for each test")
![Simple Western: Exosome Standards (HT29 cell line) [NBP3-11685] - Simple Western lane view shows lysates of exosome standards (A549 cell line), exosome standards (HT29 cell line), and COLO 205 human colorectal adenocarcinoma cell line whole cell lysate (WCL), loaded at 0.2 mg/mL. A specific band was detected for EpCAM/TROP-1 at approximately 52 kDa (as indicated) using 10 ug/mL of Goat Anti-Human EpCAM/TROP-1 Antigen Affinity-purified Polyclonal Antibody. This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/Exosome-Standards-HT29-cell-line-Simple-Western-NBP3-11685-img0002.jpg "Simple Western: Exosome Standards (HT29 cell line) [NBP3-11685] - Simple Western lane view shows lysates of exosome standards (A549 cell line), exosome standards (HT29 cell line), and COLO 205 human colorectal adenocarcinoma cell line whole cell lysate (WCL), loaded at 0.2 mg/mL. A specific band was detected for EpCAM/TROP-1 at approximately 52 kDa (as indicated) using 10 ug/mL of Goat Anti-Human EpCAM/TROP-1 Antigen Affinity-purified Polyclonal Antibody. This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.")
![Simple Western: Exosome Standards (HT29 cell line) [NBP3-11685] - Simple Western lane view shows lysates of Exosome Standards (A549 cell line), exosome standards (HT29 cell line), and COLO 205 human colorectal adenocarcinoma cell line whole cell lysate (WCL), loaded at 0.2 mg/mL. A specific band was detected for Annexin V at approximately 38 kDa (as indicated) using 50 ug/mL of Mouse Anti-Human Annexin V Monoclonal Antibody. This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/Exosome-Standards-HT29-cell-line-Simple-Western-NBP3-11685-img0003.jpg "Simple Western: Exosome Standards (HT29 cell line) [NBP3-11685] - Simple Western lane view shows lysates of Exosome Standards (A549 cell line), exosome standards (HT29 cell line), and COLO 205 human colorectal adenocarcinoma cell line whole cell lysate (WCL), loaded at 0.2 mg/mL. A specific band was detected for Annexin V at approximately 38 kDa (as indicated) using 50 ug/mL of Mouse Anti-Human Annexin V Monoclonal Antibody. This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.")
![Simple Western: Exosome Standards (HT29 cell line) [NBP3-11685] - Simple Western lane view shows lysates of Exosome Standards (A549 cell line), exosome standards (HT29 cell line), and COLO 205 human colorectal adenocarcinoma cell line whole cell lysate (WCL), loaded at 0.2 mg/mL. A specific band was detected for Flotillin-1 at approximately 51 kDa (as indicated) using 1:250 ug/mL of Flotillin-1 Antibody (NBP1-79022). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/Exosome-Standards-HT29-cell-line-Simple-Western-NBP3-11685-img0004.jpg "Simple Western: Exosome Standards (HT29 cell line) [NBP3-11685] - Simple Western lane view shows lysates of Exosome Standards (A549 cell line), exosome standards (HT29 cell line), and COLO 205 human colorectal adenocarcinoma cell line whole cell lysate (WCL), loaded at 0.2 mg/mL. A specific band was detected for Flotillin-1 at approximately 51 kDa (as indicated) using 1:250 ug/mL of Flotillin-1 Antibody (NBP1-79022). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.")
![Simple Western: Exosome Standards (HT29 cell line) [NBP3-11685] - Simple Western lane view shows lysates of Exosome Standards (A549 cell line), exosome standards (HT29 cell line), and COLO 205 human colorectal adenocarcinoma cell line whole cell lysate (WCL), loaded at 0.2 mg/mL. A specific band was detected for Alix Antibody at approximately 106 kDa (as indicated) using 1:10 ug/mL of Alix Antibody (NBP1-49701). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/Exosome-Standards-HT29-cell-line-Simple-Western-NBP3-11685-img0005.jpg "Simple Western: Exosome Standards (HT29 cell line) [NBP3-11685] - Simple Western lane view shows lysates of Exosome Standards (A549 cell line), exosome standards (HT29 cell line), and COLO 205 human colorectal adenocarcinoma cell line whole cell lysate (WCL), loaded at 0.2 mg/mL. A specific band was detected for Alix Antibody at approximately 106 kDa (as indicated) using 1:10 ug/mL of Alix Antibody (NBP1-49701). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.")
