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EpCAM/CD9 Exosome ELISA Kit

Quantification of EpCAM positive exosomes in blood samples and cell culture supernatants

This product is a two-step sandwich ELISA, which utilizes high-performance anti-EpCAM and anti-CD9 monoclonal antibodies to directly and quantitatively detect EpCAM molecules on the surface of exosomes in human blood samples and cell culture supernatants.

Human EpCAM protein is a transmembrane glycoprotein of 314 aa.
In the majority of cancer tissues, EpCAM is frequently overexpressed. Especially high abundant levels of EpCAM expression can be observed in carcinomas derived from colon, rectum, breast, lung and prostate.１） Besides affecting intercellular adhesion, EpCAM influences other important cellular functions relevant to tumor progression including cell proliferation and cancer stemness.２）

Exosomes are membrane vesicles which are secreted from many types of cells in the body and have a diameter of about 30nm~200nm. Recent reports demonstrate that most types of cancer tissue secret large numbers of exosomes that carry molecular information about the parental tumor cells.3） The information about exosomes derived from cancer cells could be a important tool for the clinical investigation.4）

However, as the analysis of exosome requires exosome purification processes using ultracentrifugation and so on, there were no rapid and convenient methods for direct measurement of exosome until now.

Kit Principle

This ELISA kit uses two-step sandwich ELISA principle. The ELISA plate provided in this kit has been pre-coated with an anti-CD9 antibody.

First, samples were added onto the plate to capture exosomes by the anti-CD9 antibody. Next, HRP conjugated anti-EpCAM antibody will be added to react with EpCAM on the surface of exosomes. Finally, substrate will be added, then measure the coloring by the plate reader to quantitate sample exosomes.

Features

  • ●Directly quantitate exosomal EpCAM in human blood samples or cell culture supernatant.
  • ●No special equipment is required. Standard microplate reader capable of reading at 450nm will do the job.
  • ●Utilize EpCAM/CD9 fusion protein (Standard Protein), instead of unstable/hard to store exosome itself, to implement stability and reproducibility.
  • ●Normalization with a standard curve using EpCAM/CD9 fusion protein (Standard Protein) enable to relative quantitate each sample.
  • ●Detect human EpCAM positive exosomes by two-step sandwich method using immobilized anti-CD9 antibody and HRP conjugated anti-EpCAM antibody.

Data

Figure 1. Standard curve

As an example, the graph of absorbance (OD450) against standard concentration is drawn as shown in Figure 1. However, draw a new standard curve for each assay to calculate the concentration in the sample.

(Plotted is the value obtained by subtracting the blank absorbance from the absorbance of each standard protein concentration)

Product List

EpCAM positive Exosome ELISA Kit

Product nameCatalog Number
EpCAM/CD9 Exosome ELISA Kit, Human HAK-HELEPCAM09-1

References

  1. 1） PT. Went, A. Lugli, S. Meier, et al.: Hum Pathol., 35, 122 (2004).
  2. 2） K. Laura, W. Stefan, P. Klaus: Cell Stress., 3, 165(2019).
  3. 3） AV. Vlassov, S. Magdaleno, R. Setterquist, et al.: Biochim Biophys Acta., 1820, 940 (2012).
  4. 4） C. Théry, S. Amigorena, G. Raposo, et al.: Curr Protoc Cell Biol., 30, Unit 3.22 (2006).

Product List

Exosome Related Antibodies

Product nameCatalog Number
Anti human CD63 Antibody, Clone A1-9A2 HAK-ANTI-CD63-A1-9A2
Anti human CD90 Antibody, Clone A6-10G9 HAK-ANTI-CD90-A6-10G9
Anti dog CD9 Antibody, Clone A1-2G10 HAK-ANTI-DCD9-2G10-1
Anti dog CD63 Antibody, Clone A2-1A4 HAK-ANTI-DCD63-1A4-1