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Human HER2 protein (extracellular domain), recombinant

Product ID: HER2 ECD (SKU#: FCL0220)

Price:

$497.00

Description

HER2 is also known as receptor tyrosine-protein kinase erbB-2 (v-erb-b2 erythroblastic leukemia viral oncogene homolog 2), metastatic lymph node gene 19 protein, proto-oncogene Neu, p185erbB2. It is a member of the epidermal growth factor receptor (EGFR) family of receptor tyrosine kinases (RTK). Four members of the EGFR family have been identified: EGFR (ERBB1, HER1), HER2 (ERBB2), HER3 (ERBB3) and HER4 (ERBB4). They typically contain an extracellular ligand binding domain (ECD), a transmembrane domain (TM), and an intracellular kinase domain that can interact with a multitude of signaling molecules and exhibit both ligand-dependent and ligand-independent activity. EGFR signaling is initiated by ligand binding to the extracellular ligand binding domain. This initiates receptor homo-/hetero-dimerization and autophosphorylation by the intracellular kinase domain, resulting in receptor activation. Following activation, phosphorylation of cytoplasmic substrates occurs and a signaling cascade is initiated that drives many cellular responses, including changes in gene expression, cytoskeletal rearrangement, anti-apoptosis and increased cell proliferation. HER2 has no ligand binding domain of its own and therefore cannot bind growth factors. However, it does bind tightly to other ligand-bound EGFR family members to form a heterodimer, stabilizing ligand binding and enhancing kinase-mediated activation of downstream signalling pathways. Amplification or overexpression of HER2 has been reported in multiple cancers, including breast and ovarian tumors. HER2 is the target of several anti-cancer therapeutics.

Gene Symbol: HER2; ERBB2; CD340; NGL; TKR1; MLN19; HER-2/neu

NCBI Gene ID: 2064

Uniprot Entry: P04626

Construct Details: The recombinant human HER2 extracellular or ecto-domain (ECD) is expressed as a 630 amino acid protein consisting of Thr23-Thr652 region of HER2 and a C-terminal poly-Histidine tag, which exists as a monomer

Source: Human cells stably expressing HER2 ECD and growing in chemical-defined media with no animal components or antibiotics.

Deduced Amino Acid Sequence:

TQVCTGTDMKLRLPASPETHLDMLRHLYQGCQVVQGNLELTYLPTNASLSFLQDIQEVQGYVLIAHNQVRQVPLQRLRIVR

GTQLFEDNYALAVLDNGDPLNNTTPVTGASPGGLRELQLRSLTEILKGGVLIQRNPQLCYQDTILWKDIFHKNNQLALTLI

DTNRSRACHPCSPMCKGSRCWGESSEDCQSLTRTVCAGGCARCKGPLPTDCCHEQCAAGCTGPKHSDCLACLHFNHSGICE

LHCPALVTYNTDTFESMPNPEGRYTFGASCVTACPYNYLSTDVGSCTLVCPLHNQEVTAEDGTQRCEKCSKPCARVCYGLG

MEHLREVRAVTSANIQEFAGCKKIFGSLAFLPESFDGDPASNTAPLQPEQLQVFETLEEITGYLYISAWPDSLPDLSVFQN

LQVIRGRILHNGAYSLTLQGLGISWLGLRSLRELGSGLALIHHNTHLCFVHTVPWDQLFRNPHQALLHTANRPEDECVGEG

LACHQLCARGHCWGPGPTQCVNCSQFLRGQECVEECRVLQGLPREYVNARHCLPCHPECQPQNGSVTCFGPEADQCVACAH

YKDPPFCVARCPSGVKPDLSYMPIWKFPDEEGACQPCPINCTHSCVDLDDKGCPAEQRASPLTSTGHHHHHHHH

M.W.: Calculated molecular mass (kDa): 70.7; Estimated by SDS-PAGE under reducing condition (kDa): ~75 (see the gel image) suggesting that the protein is glycosylated

Calculated PI: 6.01

Calculated Extinction Coefficients (M-1 cm-1, at 280nm): 65465

Purity: >95% judged by SDS-PAGE under reducing condition (see the gel image above)

Formulation: Supplied at 0.5 mg/ml in sterile PBS pH7.4 (carrier free).

Endotoxin Level: <0.1 EU per 1 μg of purified recombinant protein determined by the LAL method

Biological Activity: The recombinant protein binds anti-HER2 mAbs (SKU#MAB0061, #MAB0203, #MAB0334) with high affinity (K_D < 1 nM - see the Technical Data) and therapeutic antibodies such as Herceptin^® with high affinity (K_D < 10 nM) measured by ELISA with 10 ng of this protein coated per well.

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable. Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose. Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules. Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction. Your use of this product constitutes acceptance of the terms of this limited use agreement. Please refer to our “terms & conditions” for details. If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.

Recombinant HER2 ECD protein (SKU#FCL0220) was immobilized at 10 ng/well on a 96-well ELISA plate and incubated with serially diluted anti-HER2 human IgG1 monoclonal antibody (SKU#MAB0203). Anti-EGFR (SKU#MAB0118) human IgG1 monoclonal antibody was used as a control to demonstrate the specificity of HER2 binding under the experimental conditions. The bound antibodies were detected by goat anti-human IgG HRP-conjugated secondary antibody. The ELISA binding curve was fit and K_D was calculated for anti-HER2 : HER2 binding (affinity/avidity) to be 299 ± 13 pM.

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

The product is shipped at 4°C. Upon receipt, centrifuge the product briefly before opening the vial. It is recommended to store small aliquots at the temperature below –20°C for long-term storage and the product is stable for 3 months. The undiluted protein can be stored at 4°C for no more than 2 weeks.

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

1. Science 230: 1132–9, 1985
2. Nature 319:230-234, 1986
3. Science 229:974-976, 1985
4. Cancer Cell 5:317-328, 2004

Product datasheet (pdf) can be downloaded here: FCL0220-PDS.pdf