Recombinant Human EpCAM/TROP1 Fc Chimera Avi-tag Protein, CF

Biotinylated
Catalog # Availability Size / Price Qty
AVI960-050
Recombinant Human EpCAM/TROP1 Fc Chimera Avi-tag Protein, CF Binding Activity
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Recombinant Human EpCAM/TROP1 Fc Chimera Avi-tag Protein, CF Summary

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its binding ability in a functional ELISA. When Human EpCAM/TROP1 Affinity Purified Polyclonal Antibody (Catalog # AF960) is immobilized at 0.25 µg/mL (100 µL/well), it binds to Biotinylated Recombinant Human EpCAM/TROP‑1 Fc Chimera Avi-tag with an ED50 of 1-6 ng/mL. Measured by the ability of the immobilized protein to support the adhesion of the L Cells mouse fibroblast cell line. The ED50 for this effect is 0.7-2.8 μg/mL.
Source
Human embryonic kidney cell, HEK293-derived human EpCAM/TROP1 protein

Human EpCAM/TROP-1
(Gln24-Lys265)
Accession # P16422.2

IEGRMDHuman IgG1
(Pro100-Lys330)
Avi-tag
Accession #
N-terminal Sequence
Analysis
Gln24, inferred from deblocking reaction revealing Glu25.
Structure / Form
Disulfide-linked homodimer, biotinylated via Avi-tag
Predicted Molecular Mass
56 kDa
SDS-PAGE
55-75 kDa, under reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AVI960

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Data Images

Binding Activity Binding Activity View Larger

When Human EpCAM/TROP1 Affinity Purified Polyclonal Ab (Catalog # AF960) is immobilized at 0.25 μg/mL (100 μg/well), Biotinylated Recombinant Human EpCAM/TROP-1 Fc Chimera Avi-tag (Catalog # AVI960) binds with an ED50 of 1‑6 ng/mL.

SDS-PAGE SDS-PAGE View Larger

2 μg/lane of Biotinylated Recombinant Human EpCAM/TROP-1 Fc Chimera Avi-tag (Catalog # AVI960) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 55-75 kDa and 110-150 kDa, respectively.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: EpCAM/TROP1

Epithelial Cellular Adhesion Molecule (EpCAM), also known as KS1/4, gp40, GA733-2, 17-1A, and TROP‑1, is a transmembrane glycoprotein originally identified as a tumor-associated antigen due to its high expression level in rapidly growing epithelial tumors (1). EpCAM is one of several cell adhesion molecules (CAMs) that does not share the structural patterns found in the four major CAM families. Human EpCAM is composed of an extracellular domain (ECD) with two epidermal-growth-factor-like (EGF‑like) repeats, a single transmembrane domain, and a highly charged, short cytoplasmic domain. Within the mature ECD, human EpCAM shares 81% and 82% amino acid sequence identity with mouse and rat EpCAM respectively. During embryonic development, EpCAM is detected in fetal lung, kidney, liver, pancreas, skin, and germ cells (2). In adults, human EpCAM is detected in basolateral cell membranes of all simple, pseudo-stratified, and transitional epithelia, but is not detected in normal squamous stratified epithelia, mesenchymal tissue, muscular tissue, neuro-endocrine tissue, or lymphoid tissue (2). EpCAM expression has been found to increase in actively proliferating epithelia tissues and in human malignant neoplasias arising from squamousal epithelia (2, 3). It has been found that EpCAM is present on normal hepatic stem cells and is increased on cells with advanced cirrhosis due to the cells' proliferation through autocrine activation of Wnt signaling (4). Additionally, EpCAM may be beneficial in the early diagnosis of hepatocellular carcinoma (HCC) since HCC typically originates from a cirrhotic background (4). EpCAM has been shown function as a homophilic Ca2+ independent adhesion molecule (3). Homophilic adhesion via EpCAM requires the interaction of both EGF­like repeats, with the first EGF­like repeat mediating reciprocal interaction between EpCAM molecules on opposing cells, while the second repeat is involved in lateral interaction of EpCAM. Lateral interaction of EpCAM lead to the formation of dimers and tetramers (5). During homophilic adhesion the cytoplasmic tail of EpCAM interacts with the actin cytoskeleton via a direct association alpha ­actinin (6).

References
  1. Kloudova, K. et al. (2016) Oncotarget. 7:46120.
  2. Huang, L. et al. (2018) Int. J. Mol. Med. 42:1771.
  3. Munz, M. et al. (2009). J. Cancer Res. 69:5627.
  4. Khosla, R. et al. (2017). Stem Cells Trans. Med. 6:807.
  5. Balzar, M. et al. (2001) Mol. Cell. Biol. 21:2570.
  6. Balzar, M. et al. (1998) Mol. Cell. Biol. 18:4388.
Long Name
Epithelial Cell Adhesion Molecule
Entrez Gene IDs
4072 (Human); 17075 (Mouse); 102123042 (Cynomolgus Monkey)
Alternate Names
17-1A; 323/A3; ACSTD1; antigen identified by monoclonal AUA1; CD326 antigen; CD326; Cell surface glycoprotein Trop-1; chromosome 4, surface marker (35kD glycoprotein); DIAR5; EGP; EGP-2; EGP314; EGP40; EpCAM; epithelial cell adhesion molecule; Epithelial cell surface antigen; Epithelial glycoprotein 314; Epithelial glycoprotein; ESA; GA733-2; GA733-2EGP34; gp40; hEGP314; HNPCC8; KS 1/4 antigen; KS1/4; KSAHEA125; M1S2; M4S1; M4S1Ly74; Major gastrointestinal tumor-associated protein GA733-2; MIC18MH99; MOC31; TACST-1; TACSTD1; TROP1; TROP1CD326; Tumor-associated calcium signal transducer 1CO-17A

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