Recombinant EpCAM antibody

Catalog No. ABIN7543206

The Rabbit Chimeric anti-EpCAM antibody has been validated for FACS, IHC and ELISA. It is suitable to detect EpCAM in samples from Human.

Quick Overview for Recombinant EpCAM antibody (ABIN7543206)

Target: EpCAM (EPCAM) (Epithelial Cell Adhesion Molecule (EPCAM))

Antibody Type: Recombinant Antibody

Reactivity: Human

Host: Rabbit

Clonality: Chimeric

Conjugate: This EpCAM antibody is un-conjugated

Application: Flow Cytometry (FACS), Immunohistochemistry (IHC), ELISA

Clone: 3-17I

Product Details anti-EpCAM Antibody

Purpose: Anti-EpCAM [3-17I], Rabbit IgG, kappa

Specificity: This antibody binds human EpCAM. It also cross reacts with Cynomolgus monkey EpCAM.

Cross-Reactivity: Cynomolgus

Characteristics:

Original Species of Ab: Human

Original Format of Ab: IgG1

Purification: Protein A affinity purified

Immunogen: The original antibody was generated from a human naive scFv phage display library and later on made into a human IgG1 isotype format.

Isotype: IgG kappa

Application Details

Application Notes: The antibody specifically interacts with the natural EpCAM+Kato III cell line by flow cytometric analysis. The IgG format of the antibody bound to both human and cynomolgus EpCAM under non-reducing conditions in western blot analysis. In contrast, no binding of the antibody was observed to any reduced EpCAM antigen. Further, ELISA data demonstrated good binding of the scFv and IgG formats of the antibody to both human and cynomolgus EpCAM. The binding affinity of the IgG1 format to human and monkey EpCAM was measured by surface plasmon resonance (Kd= 1 nM and 0.93 nM, respectively). The ability of the IgG format of the antibody to induce ADCC was analyzed using three different breast cancer cell lines: MDA-MB-231, MDA-453, and BT-474. The results clearly demonstrated the antibody-induced ADCC in all three cell lines, MDA-MB-453, MDA-MB-231, and BT-474, in the presence of human PBMCs with respective estimated EC50 values of 0.08 ng/mL, 15 ng/mL, and 0.12 ng/mL. The maximum killings achieved were 75 % , 92 %, and 61 %, respectively. The ability of the IgG version of the antibody to induce complement-dependent cytotoxicity (CDC) was analyzed using two cell lines, KATO III and MT-3. The results clearly demonstrated that the antibody-induced CDC in the cell lines KATO III and MT-3 in the presence of human serum (EC50 = 0.28 ng/mL and 0.38 ng/mL for KATO III and MT-3 cells, respectively) (US8637017B2). Immunohistochemistry studies of the reactivity of the IgG2A format of the antibody were performed on a panel of normal human tissues. The antibody showed a strong reaction to sloughed cells of the esophagus sample, colon, breast, and lung cancers with epithelial origin. The antibody was conjugated to Strepsaporin. PCI of 3-17I-saporin attenuated cellular viability in three different EpCAM-positive cancer cell lines (MCF7, BxPC-3, and WiDr) as measured by MTS assay. Further, PCI of 3-17I-saporin attenuated both the proliferation and colony-forming ability of EpCAM-positive MCF7 cells (Lund et al., 2014, PMID: 24525727).

Comment:

This chimeric rabbit antibody was made using the variable domain sequences of the original Human IgG1 format, for improved compatibility with existing reagents, assays and techniques.

Restrictions: For Research Use only

Handling

Concentration: 1 mg/mL

Buffer: PBS with 0.02 % Proclin 300.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Store at 4°C for up to 3 months. For longer storage, aliquot and store at -20°C.

Target Details for EpCAM

Target: EpCAM (EPCAM) (Epithelial Cell Adhesion Molecule (EPCAM))

Alternative Name: EpCAM

Background: CD326, Ep-CAM, KS1/4 antigen, KSA, Epithelial cell adhesion molecule, Adenocarcinoma-associated antigen, Cell surface glycoprotein Trop-1, Epithelial cell surface antigen, Epithelial glycoprotein, EGP, Epithelial glycoprotein 314, 17-1A-antigen, EGP314, hEGP314, Major gastrointestinal tumor-associated protein GA733-2, Tumor-associated calcium signal transducer 1, EGP-2, EGP40, KSA, CO17-1A, GA733-2, 3-171

UniProt: P16422