HER2/CD9 Exosome ELISA Kit
This product is a two-step sandwich ELISA kit that uses high-performance antibodies against CD9 and HER2, which are exosome markers, to detect HER2 molecules expressed on the surface of exosomes secreted by cells in human blood or cell culture medium.
HER2 are frequently overexpressed in various human malignancies. Heterodimerization of these receptors results in the autophosphorylation of tyrosine residues within the cytoplasmic domain of the heterodimer and initiates signaling pathways leading to cellular proliferation and carcinogenesis.1）
Molecular targeted therapy has been established for breast cancer with HER2 overexpression and/or gene amplification, and immunohistochemical staining (IHC) is used to examine the overexpression of HER2.
As a general matter, because IHC tests are highly invasive, research on liquid biopsies using exosomes is also in progress.
Exosomes are also deeply involved in various diseases. HER2-positive exosomes secreted from HER2-positive breast cancer cells are problematic pathogenic factors that reduce the effectiveness of antibody drugs and promote the metastasis of surrounding cancer cells. 2）
Until now, it has not been to quickly and easily directly measure exosomes derived from cancer cells that highly express HER2.
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-HER2 antibody.
First, samples were added onto the plate to capture HER2 by the anti-HER2 antibody. Next, HRP conjugated anti-CD9 antibody will be added to react with CD9 on the surface of HER2positive exosomes.
Finally, substrate will be added, then measure the coloring by the plate reader to quantitate sample exosomes.
Features
- ●Directly quantitate HER2-positive exosomes 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 HER2/CD9 fusion protein (Standard Protein), instead of unstable/hard to store exosome itself, to implement stability and reproducibility.
- ●Normalization with a standard curve using HER2/CD9 fusion protein (Standard Protein) enable to relative quantitate each sample.
- ●Detect human HER2-positive exosomes by two-step sandwich method using immobilized anti-HER2 antibody and HRP conjugated anti-CD9 antibody.
Data
Figure 1. Standard curveAs 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)
Figure 2. SK-BR-3-Luc derived exosomes The culture medium of colon cancer cell line, SK-BR3-Luc, was collected, and exosomes were purified by ultracentrifugation. The purified exosomes, 31.25, 62.5, 125, 250, 500, 1000 and 2000ng/mL each, were added to the well, and measured using this kit (Fig.2).
Product List
HER2 positive Exosome ELISA Kit
| Product name | Catalog Number |
|---|---|
| HER2/CD9 Exosome ELISA Kit, Human | HAK-HELHER209-1 |
References
- 1) Y. Yarden, et al.: Nat. Rev. Mol. Cell. Biol. 2, 127 (2001)
- 2) V. Ciravolo, et al.: J Cell Physiol. 227, 658 (2012)
Product List
Exosome Related Antibodies
| Product name | Catalog 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 |