ExoELISA Complete Kit (CD81 Detection)

A sensitive ELISA assay for exosome quantitation based on the presence of CD81, a general exosome marker. Compatible with most biofluids.
  • Sensitive—detect as little as 500 µg protein equivalent
  • Flexible—compatible with all major exosome isolation methods (e.g. ExoQuick®, ultracentrifugation, ultrafiltration, and immunoaffinity capture) from human, mouse, and rat
  • Quantitative—calibrated internal standards enable quantitation of exosomes carrying CD81

Products

Overview

Overview

Sensitive, ELISA-based exosome quantitation

When you would like to use an ELISA-based method to find out how many exosomes you have and time is not limiting, choose one of SBI’s ExoELISA Complete Kits. Calibrated using NanoSight Analysis, the ExoELISA Complete Kit (CD81 Detection) enables calculation of exosome abundance based on the presence of CD81, a general exosome marker. Note that our newer ExoELISA-ULTRA Complete Kit (CD63 Detection) can provide faster exosome quantitation using significantly less sample.

  • Sensitive—detect as little as 500 µg protein equivalent
  • Flexible—compatible with all major exosome isolation methods (e.g. ExoQuick®, ultracentrifugation, ultrafiltration, and immunoaffinity capture) from human, mouse, and rat
  • Quantitative—calibrated internal standards enable quantitation of exosomes carrying CD81

How It Works

How It Works

Our ExoELISA Kits have all the reagents you need to run the ELISA—just add lysed exosome particles. The kits are compatible with exosomes isolated using most methods, including ExoQuick®, ExoQuick-TC®, or ultracentrifugation.

The ExoELISA assay uses a colorimetric, HRP activity-based readout using extra-sensitive TMB as the substrate.

The lysed exosome particles (and, thus, exosomal proteins) are directly immobilized onto the wells of the microtiter plate, and after binding, a blocking agent is added to prevent non-specific binding of the primary detection antibody, anti-CD81. Following addition of anti-CD81, a secondary antibody (goat anti-rabbit) linked to horseradish peroxidase (HRP) is also added to amplify the signal and increase assay sensitivity.

The amount of CD81 is measured via activity of the bound HRP-secondary antibody using a colorimetric assay with extra-sensitive TMB as the substrate. The accumulation of colored product is proportional to the amount of CD81 present in each well, and is measured using a microtiter plate reader at 450 nm absorbance.

Each ExoELISA Kit includes a set of standards calibrated to a known amount of exosome particles as determined by NanoSight analysis. These standards can be used to generate a calibration curve enabling quantitation of exosomes carrying CD81 from the ExoELISA data.An example ExoELISA CD81 calibration curve using the included exosome standards.The number of exosome particles in the standards supplied with each ExoELISA Kit are quantified via NanoSight Analysis.

Supporting Data

FAQs

Resources

Citations

  • Han, D, et al. (2024) Current Technology for Production, Isolation, and Quality Control of Extracellular Vesicles. Biomedical Applications of Extracellular Vesicles. 2024;:117-146. Link: Biomedical Applications of Extracellular Vesicles
  • Bhagwan Valjee, R, et al. (2024) Investigation of exosomal tetraspanin profile in sepsis patients as a promising diagnostic biomarker. Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals. 2024;:1-12. PM ID: 38354024
  • Gao, H, et al. (2024) Extracellular vesicles from organoid-derived human retinal progenitor cells prevent lipid overload-induced retinal pigment epithelium injury by regulating fatty acid metabolism. Journal of extracellular vesicles. 2024; 13(1):e12401. PM ID: 38151470
  • Byappanahalli, A, et al. (2024) Extracellular vesicle mitochondrial DNA levels are associated with race and mitochondrial DNA haplogroup. iScience. 2024; 27(1):108724. Link: iScience
  • Nagao, Y, et al. (2024) Uterine leiomyosarcoma cell-derived extracellular vesicles induce the formation of cancer-associated fibroblasts. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease. 2024;:167103. PM ID: 38417460
  • Rajeev Kumar, S, Sakthiswary, R & Lokanathan, Y. (2024) Potential Therapeutic Application and Mechanism of Action of Stem Cell-Derived Extracellular Vesicles (EVs) in Systemic Lupus Erythematosus (SLE). International journal of molecular sciences. 2024; 25(4). PM ID: 38397121
  • Huang, Y, et al. (2024) Identification of a Serum Exosome-Derived lncRNA‒miRNA‒mRNA ceRNA Network in Patients with Endometriosis. Clinical and Experimental Obstetrics & Gynecology. 2024; 51(2):51. Link: Clinical and Experimental Obstetrics & Gynecology
  • Muraoka, A, et al. (2024) Small extracellular vesicles in follicular fluids for predicting reproductive outcomes in assisted reproductive technology. Communications medicine. 2024; 4(1):33. PM ID: 38418565
  • Liang, W, Najor, RH & Gustafsson, ÅB. (2024) Protocol to separate small and large extracellular vesicles from mouse and human cardiac tissues. STAR protocols. 2024; 5(1):102914. PM ID: 38386549
  • Pallares-Rusiñol, A, et al. (2023) Advances in exosome analysis. Advances in clinical chemistry. 2023; 112:69-117. PM ID: 36642486
  • Lee, S, et al. (2023) Mesenchymal stem cell-derived extracellular vesicles subvert Th17 cells by destabilizing RORγt through posttranslational modification. Experimental & molecular medicine. 2023;. PM ID: 36964252
  • Cai, J, et al. (2023) Exosomes Derived From Kartogenin-Preconditioned Mesenchymal Stem Cells Promote Cartilage Formation and Collagen Maturation for Enthesis Regeneration in a Rat Model of Chronic Rotator Cuff Tear. The American journal of sports medicine. 2023; 51(5):1267-1276. PM ID: 36917828
  • Nguyen, CM, et al. (2023) Placental Exosomes as Biomarkers for Maternal Diseases: Current Advances in Isolation, Characterization, and Detection. ACS sensors. 2023; 8(7):2493-2513. PM ID: 37449399
  • Taha, H. (2023) Biomarkers in CNS-originating Extracellular Vesicles for Parkinson’s disease and Multiple System Atrophy. Thesis. 2023;. Link: Thesis
  • Ayala-Mar, S & Gonzalez-Valdez, J. (2023) Research and Development of Emerging Technologies for Exosome-based Cancer Diagnostics and Therapeutics. laccei.org. 2023;. Link: laccei.org
  • Rowart, P, et al. (2023) Fast and Efficient Isolation of Exosomes from Stem Cells Using a Combination of Single-Use Bioreactors, High-Speed-and Ultracentrifugation. eppendorf.com. 2023;. Link: eppendorf.com
  • Otahal, A, et al. (2023) Extracellular Vesicle Isolation and Characterization for Applications in Cartilage Tissue Engineering and Osteoarthritis Therapy. Methods in molecular biology (Clifton, N.J.). 2023; 2598:123-140. PM ID: 36355289
  • Guo, Q, et al. (2023) Glioblastoma upregulates SUMOylation of hnRNP A2/B1 to eliminate the tumor suppressor miR-204-3p, accelerating angiogenesis under hypoxia. Cell death & disease. 2023; 14(2):147. PM ID: 36810326
  • von Stade, DP, et al. (2023) Exosome cell origin affects in vitro markers of tendon repair in ovine macrophages and tenocytes. Tissue engineering. Part A. 2023;. PM ID: 36792933
  • Pradhan, A, et al. (2023) Association of exosomal miR-96-5p and miR-146a-5p with the disease severity in dengue virus infection. Journal of medical virology. 2023; 95(3):e28614. PM ID: 36840403

Products

Overview

Overview

Sensitive, ELISA-based exosome quantitation

When you would like to use an ELISA-based method to find out how many exosomes you have and time is not limiting, choose one of SBI’s ExoELISA Complete Kits. Calibrated using NanoSight Analysis, the ExoELISA Complete Kit (CD81 Detection) enables calculation of exosome abundance based on the presence of CD81, a general exosome marker. Note that our newer ExoELISA-ULTRA Complete Kit (CD63 Detection) can provide faster exosome quantitation using significantly less sample.

  • Sensitive—detect as little as 500 µg protein equivalent
  • Flexible—compatible with all major exosome isolation methods (e.g. ExoQuick®, ultracentrifugation, ultrafiltration, and immunoaffinity capture) from human, mouse, and rat
  • Quantitative—calibrated internal standards enable quantitation of exosomes carrying CD81

How It Works

How It Works

Our ExoELISA Kits have all the reagents you need to run the ELISA—just add lysed exosome particles. The kits are compatible with exosomes isolated using most methods, including ExoQuick®, ExoQuick-TC®, or ultracentrifugation.

The ExoELISA assay uses a colorimetric, HRP activity-based readout using extra-sensitive TMB as the substrate.

The lysed exosome particles (and, thus, exosomal proteins) are directly immobilized onto the wells of the microtiter plate, and after binding, a blocking agent is added to prevent non-specific binding of the primary detection antibody, anti-CD81. Following addition of anti-CD81, a secondary antibody (goat anti-rabbit) linked to horseradish peroxidase (HRP) is also added to amplify the signal and increase assay sensitivity.

The amount of CD81 is measured via activity of the bound HRP-secondary antibody using a colorimetric assay with extra-sensitive TMB as the substrate. The accumulation of colored product is proportional to the amount of CD81 present in each well, and is measured using a microtiter plate reader at 450 nm absorbance.

Each ExoELISA Kit includes a set of standards calibrated to a known amount of exosome particles as determined by NanoSight analysis. These standards can be used to generate a calibration curve enabling quantitation of exosomes carrying CD81 from the ExoELISA data.An example ExoELISA CD81 calibration curve using the included exosome standards.The number of exosome particles in the standards supplied with each ExoELISA Kit are quantified via NanoSight Analysis.

Supporting Data

FAQs

Citations

  • Han, D, et al. (2024) Current Technology for Production, Isolation, and Quality Control of Extracellular Vesicles. Biomedical Applications of Extracellular Vesicles. 2024;:117-146. Link: Biomedical Applications of Extracellular Vesicles
  • Bhagwan Valjee, R, et al. (2024) Investigation of exosomal tetraspanin profile in sepsis patients as a promising diagnostic biomarker. Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals. 2024;:1-12. PM ID: 38354024
  • Gao, H, et al. (2024) Extracellular vesicles from organoid-derived human retinal progenitor cells prevent lipid overload-induced retinal pigment epithelium injury by regulating fatty acid metabolism. Journal of extracellular vesicles. 2024; 13(1):e12401. PM ID: 38151470
  • Byappanahalli, A, et al. (2024) Extracellular vesicle mitochondrial DNA levels are associated with race and mitochondrial DNA haplogroup. iScience. 2024; 27(1):108724. Link: iScience
  • Nagao, Y, et al. (2024) Uterine leiomyosarcoma cell-derived extracellular vesicles induce the formation of cancer-associated fibroblasts. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease. 2024;:167103. PM ID: 38417460
  • Rajeev Kumar, S, Sakthiswary, R & Lokanathan, Y. (2024) Potential Therapeutic Application and Mechanism of Action of Stem Cell-Derived Extracellular Vesicles (EVs) in Systemic Lupus Erythematosus (SLE). International journal of molecular sciences. 2024; 25(4). PM ID: 38397121
  • Huang, Y, et al. (2024) Identification of a Serum Exosome-Derived lncRNA‒miRNA‒mRNA ceRNA Network in Patients with Endometriosis. Clinical and Experimental Obstetrics & Gynecology. 2024; 51(2):51. Link: Clinical and Experimental Obstetrics & Gynecology
  • Muraoka, A, et al. (2024) Small extracellular vesicles in follicular fluids for predicting reproductive outcomes in assisted reproductive technology. Communications medicine. 2024; 4(1):33. PM ID: 38418565
  • Liang, W, Najor, RH & Gustafsson, ÅB. (2024) Protocol to separate small and large extracellular vesicles from mouse and human cardiac tissues. STAR protocols. 2024; 5(1):102914. PM ID: 38386549
  • Pallares-Rusiñol, A, et al. (2023) Advances in exosome analysis. Advances in clinical chemistry. 2023; 112:69-117. PM ID: 36642486
  • Lee, S, et al. (2023) Mesenchymal stem cell-derived extracellular vesicles subvert Th17 cells by destabilizing RORγt through posttranslational modification. Experimental & molecular medicine. 2023;. PM ID: 36964252
  • Cai, J, et al. (2023) Exosomes Derived From Kartogenin-Preconditioned Mesenchymal Stem Cells Promote Cartilage Formation and Collagen Maturation for Enthesis Regeneration in a Rat Model of Chronic Rotator Cuff Tear. The American journal of sports medicine. 2023; 51(5):1267-1276. PM ID: 36917828
  • Nguyen, CM, et al. (2023) Placental Exosomes as Biomarkers for Maternal Diseases: Current Advances in Isolation, Characterization, and Detection. ACS sensors. 2023; 8(7):2493-2513. PM ID: 37449399
  • Taha, H. (2023) Biomarkers in CNS-originating Extracellular Vesicles for Parkinson’s disease and Multiple System Atrophy. Thesis. 2023;. Link: Thesis
  • Ayala-Mar, S & Gonzalez-Valdez, J. (2023) Research and Development of Emerging Technologies for Exosome-based Cancer Diagnostics and Therapeutics. laccei.org. 2023;. Link: laccei.org
  • Rowart, P, et al. (2023) Fast and Efficient Isolation of Exosomes from Stem Cells Using a Combination of Single-Use Bioreactors, High-Speed-and Ultracentrifugation. eppendorf.com. 2023;. Link: eppendorf.com
  • Otahal, A, et al. (2023) Extracellular Vesicle Isolation and Characterization for Applications in Cartilage Tissue Engineering and Osteoarthritis Therapy. Methods in molecular biology (Clifton, N.J.). 2023; 2598:123-140. PM ID: 36355289
  • Guo, Q, et al. (2023) Glioblastoma upregulates SUMOylation of hnRNP A2/B1 to eliminate the tumor suppressor miR-204-3p, accelerating angiogenesis under hypoxia. Cell death & disease. 2023; 14(2):147. PM ID: 36810326
  • von Stade, DP, et al. (2023) Exosome cell origin affects in vitro markers of tendon repair in ovine macrophages and tenocytes. Tissue engineering. Part A. 2023;. PM ID: 36792933
  • Pradhan, A, et al. (2023) Association of exosomal miR-96-5p and miR-146a-5p with the disease severity in dengue virus infection. Journal of medical virology. 2023; 95(3):e28614. PM ID: 36840403