Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total E-Cadherin Sandwich ELISA Kit #7886

When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

Important Ordering Details: Product is assembled upon order to ensure maximum activity. Domestic: Please allow up to two weeks for your order to be processed and shipped. International: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.

Kit Includes Volume Solution Color
E-Cadherin Mouse Antibody Coated Microwells 96 tests
E-Cadherin Rabbit Detection Antibody 11 ml Green
Anti-rabbit IgG, HRP-linked Antibody 11 ml Red
TMB Substrate #7004 11 ml Colorless
STOP Solution #7002 11 ml Colorless
Sealing Tape 2 sheets
ELISA Wash Buffer (20X) 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
Cell Lysis Buffer (10X) #9803 15 ml Yellowish

Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer (10X), which is stored at –20°C (packaged separately).

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Description

The PathScan® Total E-Cadherin Sandwich ELISA Kit #7886 is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of E-cadherin. An E-cadherin mouse antibody has been coated onto the microwells. After incubation with cell lysates, E-cadherin is captured by the coated antibody. Following extensive washing, an E-Cadherin rabbit detection antibody is added to detect the captured E-cadherin. Anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of E-cadherin.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

CST's PathScan® Total E-Cadherin Sandwich ELISA Kit #7886 detects endogenous levels of E-cadherin. As shown in Figure 1, using the Total E-Cadherin Sandwich ELISA Kit #7886, a high level of E-cadherin is detected in A431 and MCF-7 cells, but not in E-cadherin null Jurkat cells. The PathScan® Total E-Cadherin Sandwich ELISA Kit #7886 does not cross-react with human N-, P- or VE-cadherin. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

ELISA - Western correlation

ELISA - Western correlation

Figure 1. E-cadherin is detected by PathScan® Total E-Cadherin Sandwich ELISA Kit #7886. Lysates were prepared from various cell lines and analyzed for E-cadherin using PathScan® Total E-Cadherin Sandwich ELISA Kit #7886 or by western blotting. Absorbance readings at 450 nm are shown in the top figure while the corresponding western blot using an E-cadherin antibody is shown in the bottom figure.

Sensitivity

Sensitivity

Figure 2. The relationship between the protein concentration of the lysate from A431 and Jurkat cells and the absorbance at 450 nm is shown.

Background

Cadherins are a superfamily of transmembrane glycoproteins that contain cadherin repeats of approximately 100 residues in their extracellular domain. Cadherins mediate calcium-dependent cell-cell adhesion and play critical roles in normal tissue development (1). The classic cadherin subfamily includes N-, P-, R-, B- and E-cadherins as well as about ten other members that are found in adherens junctions, a cellular structure near the apical surface of polarized epithelial cells. The cytoplasmic domain of classical cadherins interacts with β-catenin, γ-catenin (also called plakoglobin), and p120 catenin. β-catenin and γ-catenin associate with α-catenin, which links the cadherin-catenin complex to the actin cytoskeleton (1,2). While β- and γ-catenin play structural roles in the junctional complex, p120 regulates cadherin adhesive activity and trafficking (1-4). E-cadherin is considered an active suppressor of invasion and growth of many epithelial cancers (1-3). Recent studies indicate that cancer cells have up-regulated N-cadherin in addition to loss of E-cadherin. This change in cadherin expression is called the "cadherin switch". N-cadherin cooperates with the FGF receptor, leading to overexpression of MMP-9 and cellular invasion (3). In endothelial cells, VE-cadherin signaling, expression, and localization correlate with vascular permeability and tumor angiogenesis (5,6). Expression of P-cadherin, which is normally present in epithelial cells, is also altered in ovarian and other human cancers (7,8).

  1. Wheelock, M.J. and Johnson, K.R. (2003) Annu. Rev. Cell. Dev. Biol. 19, 207-235.
  2. Christofori, G. (2003) EMBO J. 22, 2318-2323.
  3. Hazan, R.B. et al. (2004) Ann. NY Acad. Sci. 1014, 155-163.
  4. Bryant, D.M. and Stow, J.L. (2004) Trends Cell Biol. 14, 427-434.
  5. Rabascio, C. et al. (2004) Cancer Res. 64, 4373-4377.
  6. Yamaoka-Tojo, M. et al. (2006) Arterioscler. Thromb. Vasc. Biol. 26, 1991-1997.
  7. Patel, I.S. et al. (2003) Int. J. Cancer 106, 172-177.
  8. Sanders, D.S. et al. (2000) J. Pathol. 190, 526-530.

Application References

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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