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Encompassing Amino Acids: 23-652
Application: Useful for the study of screening inhibitors, and selectivity profiling.
Assay Conditions: HER2 protein was used to coat a 96-well plate overnight at 4°C (50 µl/well at a concentration of 100 ng/well in PBS). The next day the plate was washed 3 times with PBST and blocked using 200 µl of Blocking Buffer 3 (#79743) for 2 hours at Room Temperature (RT). After removing the blocking buffer, 50 µl/well of purified Anti-Her2 Antibody (#101689), serially diluted in 1x PP-02 Buffer, was added for 1 hour at RT. The plate was washed and incubated for 60 minutes with an Anti-Fc-HRP-labeled antibody 1:1000 in Blocking Buffer 3 (50 µl/well) and washed again. Finally, 100 µl ECL substrate was added to each well and the plate was read immediately in a luminometer or microtiter-plate reader capable of reading chemiluminescence.
Background: HER2 (human epidermal growth factor receptor 2), also known as erbB-2 or CD340, is a tyrosine kinase of the EFGR family of proteins. There is no known ligand, but it can form homodimers or heterodimers with other HER proteins. Once active, it activates the MAPK (mitogen-activated protein kinase) and PI3K (phosphatidylinositol-3 kinase) signaling pathways resulting in cell cycle progression and cell proliferation. HER2 over-expression is also known to occur in breast, ovarian, stomach, lung adenocarcinoma, aggressive forms of uterine cancer and gastric cancer. In 1990 the FDA approved the use of the monoclonal antibody trastuzumab in breast and stomach cancer. Other strategies to target HER2 that have been approved include ADCs (antibody-drug conjugate) and margetuximab (an HER2 antibody that can alter the Fc-receptor affinity to CD16 and induce cytotoxicity). The development of treatments able to target early-stage cancer, with minimal side effects and resistance development, will bring major benefits to HER2+ oncology patients.
Description: Recombinant human HER2 (human epidermal growth factor receptor 2), encompassing amino acids 23-652 (extracellular domain). This construct contains an Avi-Tag™ followed by an His-tag (6xHis). This protein was affinity purified.
Format: Aqueous buffer solution.
Formulation: 8 mM phosphate, pH 7.4, 110 mM NaCl, 2.2 mM KCl, and 20% glycerol
Genbank: NM_004448.4
Purity: ≥90%
Supplied As: At least 6 months at -80°C.
Target: C-terminal Avi-His-Tag
Uniprot: P04626
Warnings: Avoid multiple freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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