SmartSEC-DeLipo™ Advanced sEV Isolation Kit

Purer EVs with Less Lipoproteins

Elevate your small extracellular vesicle (sEV) isolation game with our cutting-edge SmartSEC-DeLipoTM kit, engineered to provide unmatched purity and quality while efficiently depleting lipoprotein contaminants. Harnessing the power of SmartSECTM coupled with innovative lipoprotein removal technology, this kit streamlines the sEV isolation process, ensuring pure sEVs samples for downstream analyses.
  • New generation of mixed-mode chromatography
  • Advanced Lipoprotein Depletion
  • Fast and Easy Isolation of High Yield and Purity sEV
  • Versatile Compatibility with Various Applications

Products

Catalog Number Description Size Price Quantity Add to Cart
SSEC-DLP-200A SmartSEC-DeLipo Single EV Isolation System 10 Reactions $498
- +
DLP-20A DeLipo lipoprotein removal column 20 Reactions $298
- +
SSEC-DLP-096A SmartSEC-DeLipo HT EV Isolation System 96 Reactions $2945
- +

Overview

Overview

Importance of Removing Lipoproteins from sEV

Extracellular vesicles (EVs) from blood are of great importance to understand the biological role of circulating EVs and to develop EVs as biomarkers of disease. However, isolating EVs from blood poses challenges due to the simultaneous presence of lipoprotein particles. Lipoproteins may interfere with downstream analyses or functional assays performed on EVs. For instance, lipoproteins contain microRNAs as their cargos. In studies investigating the biological roles of EVs or their potential as biomarkers, the presence of lipoproteins could confound the results or mask the true effects of EVs. Recently, several studies showed that ultracentrifugation or size exclusion chromatography (SEC) alone cannot completely separate blood EVs from lipoprotein particles. Therefore, improved separation of EVs from lipoproteins is crucial for a detailed functional analysis of circulating EVs, thus making blood a viable source for EV biomarker discovery.

Advanced sEV Isolation with SmartSEC-DeLipo

Our cutting-edge SmartSEC-DeLipoTM kit was engineered to provide unmatched purity and quality while efficiently depleting lipoprotein contaminants for sEV isolation. Harnessing the power of SmartSEC coupled with innovative lipoprotein removal technology, this kit streamlines the sEV isolation process, ensuring much purer sEVs samples for downstream analyses.

  • New generation of mixed-mode chromatography: Our SmartSEC-DelipoTM Kit utilizes SmartSECTM columns, a new generation of mixed-mode chromatography, which provides the dual functionality of size exclusion and affinity interaction modes. Combining the two modes within a single resin media eliminates the time consuming and labor-intensive processes of classical SEC, delivering highly purified sEV samples.
  • Advanced Lipoprotein Depletion: Selectively removes lipoproteins from the sample, eliminating interference and ensuring the isolation of pure sEV populations, experiencing a new standard of purity.
  • Efficient Workflow: With a user-friendly protocol designed for efficiency and reproducibility, researchers can easily integrate SmartSEC-DeLipoTM into their workflows, saving time and resources.
  • High Yield and Purity: Achieve consistently high yields of pure sEVs, free from lipoprotein contaminants, enabling robust downstream applications such as RNA sequencing, proteomics, and functional studies.
  • Versatile Compatibility: Compatible with various sample types containing lipoprotein including plasma, serum, breast milk et al, ensuring versatility and flexibility in experimental design.
  • Superior Performance: SmartSEC-DeLipoTM ensures consistent and reliable performance, empowering researchers with confidence in their results and facilitating reproducible experiments.
  • Various Applications: Ideal for a wide range of research applications including biomarker discovery, disease diagnostics, therapeutic development, and elucidating the role of sEVs in intercellular communication.

Experience the future of sEV isolation with SmartSEC-DeLipoTM Advanced sEV Isolation Kit. Unlock unparalleled purity, efficiency, and reliability, and accelerate your journey towards groundbreaking discoveries in the dynamic field of extracellular vesicle biology.

How It Works

The SmartSEC-DeLipo™ workflow is fast and easy. Simply apply 250 µL of cleared serum or plasma with additional column buffer or up to 0.5 mL of other biofluids directly to the pre-washed column, incubate, and centrifuge to elute the Evs then go through DeLipo column to remove lipoprotein from the isolated sEVs.

Workflow of SmartSEC-DeLipo

Figure 1. Workflow of SmartSEC-DeLipoTM

Supporting Data

SmartSEC-DeLipoTM isolates much purer sEV

SmartSEC-DeLipo™ isolates much purer sEV

Figure 2. sEV isolated with SmartSEC-DeLipoTM is much purer. (A) sEV isolated from 250ul of serum with SmartSEC-DeLipoTM shows higher expression of EV markers, such as CD9, CD81 and TSG101 than those isolated with SmartSECTM single when same amount of EV protein were applied for western blot. When same volume of eluted sEV samples were applied for western blot, EV markers’ expression is about the same for sEV isolated from 250ul of serum with SmartSEC-DeLipoTM or SmartSECTM single. (B) sEV isolated from 250ul of plasma or breast milk with SmartSEC-DeLipoTM shows higher expression of EV markers, such as CD9, CD81 and TSG101 than those isolated with SmartSECTM single when same amount of EV proteins was applied for western blot. (C) fNTA data shows that sEV particle number were decreased in plasma or breast milk sample when isolated with SmartSEC-DeLipoTM in comparison with that isolated with SmartSECTM single. However, when purity was evaluated with EV particle number per same amount of EV protein, sEV isolated with SmartSEC-DeLipoTM demonstrates higher purity than those isolated with SmartSECTM single.


SmartSEC-DeLipo removes most of the lipoproteins

SmartSEC-DeLipo removes most of the lipoproteins

Figure 3. Most of the Lipoproteins are removed in sEV isolated with SmartSEC-DeLipoTM. (A) ApoB and ApoE lipoproteins’ concentrations were evaluated in serum sample, sEV isolated from serum with SmartSECTM single and sEV isolated from serum with SmartSEC-DeLipoTM by ELISA. SmarSECTM single dramatically reduced Lipoproteins in isolated sEV and SmartSEC-DeLipo further deplete the remaining Lipoproteins in yield sEV. (B) Percentage of ApoB and ApoE depletion was calculated based on ApoB and ApoE concentration in sEV isolated with SmartSEC-DeLipoTM vs. SmartSECTM single from serum, plasma and breast milk samples.


SmartSEC-DeLipoTM isolated sEV possess typical sEV morphology

SmartSEC-DeLipo™ isolated sEV possess typical sEV morphology

Figure 4. sEV isolated with SmartSEC-DeLipoTM shows typical EV morphology. Transmission electron microscopy (TEM) of sEVs isolated from serum using SmartSEC-DeLipoTM possess typical EV morphology—intact vesicles with a double layer of membranes.


SmartSEC-DeLipoTM isolate sEV is biological functional

SmartSEC-DeLipo™ isolate sEV is biological functional

Figure 5. sEV isolated with SmartSEC-DeLipoTM is biologically functional to delivery small RNA cargo to the recipient cells. (A) Cy3-labeled control siRNA were loaded without or with sEV isolated from serum with SmartSECTM single or SmartSEC-DeLipoTM using Exo-Fect siRNA/miRNA reagent (EXFT200A-1) and delivered to HEK 293 cells. Cells imaged 36-48 hours post transfection. Shown are the bright-field images (bottom row), and fluorescence images (top row). Both sEV isolated with SmartSEC single or SmartSEC-DeLipoTM can efficiently transfer Cy3-labeled siRNA to most of the imaged cells. (B) HPRT siRNA were loaded without or with sEV isolated from serum with SmartSECTM single or SmartSEC-DeLipoTM using Exo-Fect siRNA/miRNA reagent (EXFT200A-1) and delivered to HEK 293 cells. Cells were lysed and applied for western blot 48 hours post transfection. Both sEV isolated with SmartSECTM single or SmartSEC-DeLipoTM can efficiently transfer HPRT siRNA to HEK 293 cells and achieve knockdown of the endogenous HPRT expression.


SmartSEC-DeLipoTM isolate sEV can be used for biomarker discovery

SmartSEC-DeLipo™ isolate sEV can be used for biomarker discovery

Figure 6. sEV isolated with SmartSEC-DeLipoTM can be used for small RNA profiling and biomarker discovery. sEV isolated from normal vs. pancreatic cancer patient plasma with SmartSEC-DeLipoTM were used for sEV RNA isolated using EVery100B-1. Then RNA samples were analyzed by Agilent Bioanalyzer. (A) Electrophoretic spectrums and gel images of the sEV RNA derived from the plasma of pooled normal plasma. Then sEV RNA samples from normal and pancreatic patient sample were reverse transcribed into cDNA using Every 200B-1, followed with Every miRNome Profiler (EVery600B-1) (B) Heat map of normal vs. pancreatic cancer plasma miRNA expression profiler. Representative data normalized to global mean of each sample. Pancreatic cancer patient plasma showed distinct miRNA expression profiles compared to normal plasma. (C) Differential expression of selected miRNAs in normal vs. pancreatic cancer plasma. Data normalized with global mean. Expression of miR-25-3p, miR-19b-3p, miR-451a, miR-486-5p, miR-103a-3p and miR-19a-3p are significantly upregulated, while expression of miR-185-3p, miR-424-5p, miR-140-5p and miR-125a-5p is significantly downregulated in pancreatic cancer plasma compared to normal one.


SmartSEC-DeLipoTM isolate sEV can be used for proteomic study

SmartSEC-DeLipo™ isolate sEV can be used for proteomic study

Figure 7. sEV isolated with SmartSEC-DeLipoTM can be used for proteomic studies. sEV isolated from normal vs. pancreatic cancer patient plasma with SmartSEC-DeLipoTM or SmartSECTM single were analyzed by mass spectrometry. (A) Heatmap of normal vs. pancreatic cancer plasma sEV proteomic profiler using either SmartSECTM single or SmartSEC-DeLipoTM. Pancreatic cancer patient plasma sEV showed distinct proteome compared to normal plasma. (B) sEV from normal pooled plasma isolated with either SmartSECTM or SmartSEC-DeLipoTM also showed differentiated proteome.

FAQs

Resources

Citations

Products

Catalog Number Description Size Price Quantity Add to Cart
SSEC-DLP-200A SmartSEC-DeLipo Single EV Isolation System 10 Reactions $498
- +
DLP-20A DeLipo lipoprotein removal column 20 Reactions $298
- +
SSEC-DLP-096A SmartSEC-DeLipo HT EV Isolation System 96 Reactions $2945
- +

Overview

Overview

Importance of Removing Lipoproteins from sEV

Extracellular vesicles (EVs) from blood are of great importance to understand the biological role of circulating EVs and to develop EVs as biomarkers of disease. However, isolating EVs from blood poses challenges due to the simultaneous presence of lipoprotein particles. Lipoproteins may interfere with downstream analyses or functional assays performed on EVs. For instance, lipoproteins contain microRNAs as their cargos. In studies investigating the biological roles of EVs or their potential as biomarkers, the presence of lipoproteins could confound the results or mask the true effects of EVs. Recently, several studies showed that ultracentrifugation or size exclusion chromatography (SEC) alone cannot completely separate blood EVs from lipoprotein particles. Therefore, improved separation of EVs from lipoproteins is crucial for a detailed functional analysis of circulating EVs, thus making blood a viable source for EV biomarker discovery.

Advanced sEV Isolation with SmartSEC-DeLipo

Our cutting-edge SmartSEC-DeLipoTM kit was engineered to provide unmatched purity and quality while efficiently depleting lipoprotein contaminants for sEV isolation. Harnessing the power of SmartSEC coupled with innovative lipoprotein removal technology, this kit streamlines the sEV isolation process, ensuring much purer sEVs samples for downstream analyses.

  • New generation of mixed-mode chromatography: Our SmartSEC-DelipoTM Kit utilizes SmartSECTM columns, a new generation of mixed-mode chromatography, which provides the dual functionality of size exclusion and affinity interaction modes. Combining the two modes within a single resin media eliminates the time consuming and labor-intensive processes of classical SEC, delivering highly purified sEV samples.
  • Advanced Lipoprotein Depletion: Selectively removes lipoproteins from the sample, eliminating interference and ensuring the isolation of pure sEV populations, experiencing a new standard of purity.
  • Efficient Workflow: With a user-friendly protocol designed for efficiency and reproducibility, researchers can easily integrate SmartSEC-DeLipoTM into their workflows, saving time and resources.
  • High Yield and Purity: Achieve consistently high yields of pure sEVs, free from lipoprotein contaminants, enabling robust downstream applications such as RNA sequencing, proteomics, and functional studies.
  • Versatile Compatibility: Compatible with various sample types containing lipoprotein including plasma, serum, breast milk et al, ensuring versatility and flexibility in experimental design.
  • Superior Performance: SmartSEC-DeLipoTM ensures consistent and reliable performance, empowering researchers with confidence in their results and facilitating reproducible experiments.
  • Various Applications: Ideal for a wide range of research applications including biomarker discovery, disease diagnostics, therapeutic development, and elucidating the role of sEVs in intercellular communication.

Experience the future of sEV isolation with SmartSEC-DeLipoTM Advanced sEV Isolation Kit. Unlock unparalleled purity, efficiency, and reliability, and accelerate your journey towards groundbreaking discoveries in the dynamic field of extracellular vesicle biology.

How It Works

The SmartSEC-DeLipo™ workflow is fast and easy. Simply apply 250 µL of cleared serum or plasma with additional column buffer or up to 0.5 mL of other biofluids directly to the pre-washed column, incubate, and centrifuge to elute the Evs then go through DeLipo column to remove lipoprotein from the isolated sEVs.

Workflow of SmartSEC-DeLipo

Figure 1. Workflow of SmartSEC-DeLipoTM

Supporting Data

SmartSEC-DeLipoTM isolates much purer sEV

SmartSEC-DeLipo™ isolates much purer sEV

Figure 2. sEV isolated with SmartSEC-DeLipoTM is much purer. (A) sEV isolated from 250ul of serum with SmartSEC-DeLipoTM shows higher expression of EV markers, such as CD9, CD81 and TSG101 than those isolated with SmartSECTM single when same amount of EV protein were applied for western blot. When same volume of eluted sEV samples were applied for western blot, EV markers’ expression is about the same for sEV isolated from 250ul of serum with SmartSEC-DeLipoTM or SmartSECTM single. (B) sEV isolated from 250ul of plasma or breast milk with SmartSEC-DeLipoTM shows higher expression of EV markers, such as CD9, CD81 and TSG101 than those isolated with SmartSECTM single when same amount of EV proteins was applied for western blot. (C) fNTA data shows that sEV particle number were decreased in plasma or breast milk sample when isolated with SmartSEC-DeLipoTM in comparison with that isolated with SmartSECTM single. However, when purity was evaluated with EV particle number per same amount of EV protein, sEV isolated with SmartSEC-DeLipoTM demonstrates higher purity than those isolated with SmartSECTM single.


SmartSEC-DeLipo removes most of the lipoproteins

SmartSEC-DeLipo removes most of the lipoproteins

Figure 3. Most of the Lipoproteins are removed in sEV isolated with SmartSEC-DeLipoTM. (A) ApoB and ApoE lipoproteins’ concentrations were evaluated in serum sample, sEV isolated from serum with SmartSECTM single and sEV isolated from serum with SmartSEC-DeLipoTM by ELISA. SmarSECTM single dramatically reduced Lipoproteins in isolated sEV and SmartSEC-DeLipo further deplete the remaining Lipoproteins in yield sEV. (B) Percentage of ApoB and ApoE depletion was calculated based on ApoB and ApoE concentration in sEV isolated with SmartSEC-DeLipoTM vs. SmartSECTM single from serum, plasma and breast milk samples.


SmartSEC-DeLipoTM isolated sEV possess typical sEV morphology

SmartSEC-DeLipo™ isolated sEV possess typical sEV morphology

Figure 4. sEV isolated with SmartSEC-DeLipoTM shows typical EV morphology. Transmission electron microscopy (TEM) of sEVs isolated from serum using SmartSEC-DeLipoTM possess typical EV morphology—intact vesicles with a double layer of membranes.


SmartSEC-DeLipoTM isolate sEV is biological functional

SmartSEC-DeLipo™ isolate sEV is biological functional

Figure 5. sEV isolated with SmartSEC-DeLipoTM is biologically functional to delivery small RNA cargo to the recipient cells. (A) Cy3-labeled control siRNA were loaded without or with sEV isolated from serum with SmartSECTM single or SmartSEC-DeLipoTM using Exo-Fect siRNA/miRNA reagent (EXFT200A-1) and delivered to HEK 293 cells. Cells imaged 36-48 hours post transfection. Shown are the bright-field images (bottom row), and fluorescence images (top row). Both sEV isolated with SmartSEC single or SmartSEC-DeLipoTM can efficiently transfer Cy3-labeled siRNA to most of the imaged cells. (B) HPRT siRNA were loaded without or with sEV isolated from serum with SmartSECTM single or SmartSEC-DeLipoTM using Exo-Fect siRNA/miRNA reagent (EXFT200A-1) and delivered to HEK 293 cells. Cells were lysed and applied for western blot 48 hours post transfection. Both sEV isolated with SmartSECTM single or SmartSEC-DeLipoTM can efficiently transfer HPRT siRNA to HEK 293 cells and achieve knockdown of the endogenous HPRT expression.


SmartSEC-DeLipoTM isolate sEV can be used for biomarker discovery

SmartSEC-DeLipo™ isolate sEV can be used for biomarker discovery

Figure 6. sEV isolated with SmartSEC-DeLipoTM can be used for small RNA profiling and biomarker discovery. sEV isolated from normal vs. pancreatic cancer patient plasma with SmartSEC-DeLipoTM were used for sEV RNA isolated using EVery100B-1. Then RNA samples were analyzed by Agilent Bioanalyzer. (A) Electrophoretic spectrums and gel images of the sEV RNA derived from the plasma of pooled normal plasma. Then sEV RNA samples from normal and pancreatic patient sample were reverse transcribed into cDNA using Every 200B-1, followed with Every miRNome Profiler (EVery600B-1) (B) Heat map of normal vs. pancreatic cancer plasma miRNA expression profiler. Representative data normalized to global mean of each sample. Pancreatic cancer patient plasma showed distinct miRNA expression profiles compared to normal plasma. (C) Differential expression of selected miRNAs in normal vs. pancreatic cancer plasma. Data normalized with global mean. Expression of miR-25-3p, miR-19b-3p, miR-451a, miR-486-5p, miR-103a-3p and miR-19a-3p are significantly upregulated, while expression of miR-185-3p, miR-424-5p, miR-140-5p and miR-125a-5p is significantly downregulated in pancreatic cancer plasma compared to normal one.


SmartSEC-DeLipoTM isolate sEV can be used for proteomic study

SmartSEC-DeLipo™ isolate sEV can be used for proteomic study

Figure 7. sEV isolated with SmartSEC-DeLipoTM can be used for proteomic studies. sEV isolated from normal vs. pancreatic cancer patient plasma with SmartSEC-DeLipoTM or SmartSECTM single were analyzed by mass spectrometry. (A) Heatmap of normal vs. pancreatic cancer plasma sEV proteomic profiler using either SmartSECTM single or SmartSEC-DeLipoTM. Pancreatic cancer patient plasma sEV showed distinct proteome compared to normal plasma. (B) sEV from normal pooled plasma isolated with either SmartSECTM or SmartSEC-DeLipoTM also showed differentiated proteome.

FAQs

Resources

Citations