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Encompassing Amino Acids: 26-651
Application: Useful for binding studies.
Assay Conditions: The NRG1β Protein, Human (HEK293) was coated onto a 96-well plate overnight at 4°C (25 ng/well). 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 HER4 (ErbB4), Fc Fusion (IgG1), Avi-Tag, Biotin-Labeled Recombinant (#102239), serially diluted in 1x PP-02 Buffer, was added for 1 hour at RT. After washing, the plate was incubated for 1 hour with HRP-labeled anti-FC antibody diluted 1:1000 in Blocking Buffer 3 (50 µl/well) and washed again. Finally, 100 µl of ECL substrate was added to each well and chemiluminescence was read immediately.
Background: HER4, also known as ErbB4 or receptor-tyrosine kinase 4, is a receptor tyrosine kinase protein of the EGFR family. It is required for the development of the heart, nervous system, and mammary gland, and can also be found in the postsynaptic side of neuromuscular junctions and epidermis. HER4 is activated by neuroregulin 2-4, betacellulin and heparin binding EGF-like growth factor (HB-EGF) and is involved in mitogenesis and cell differentiation. HER4 undergoes a conformational change upon ligand binding and forms homodimers or heterodimers with other HER proteins to activate the MAPK (mitogen-activated protein kinases) and PI3K (phosphoinositide 3-kinase)/AKT pathways. Mutations in this protein can result in cancer but have also been linked to ALS (amyotrophic lateral sclerosis) type 19 and schizophrenia. The role of this protein as oncogene or tumor suppressor is still not fully determined, potentially being dependent on the cancer type. Several HER4 inhibitors have been developed and are undergoing clinical trials. Afatinib, a pan-HER inhibitor, has been approved for the treatment of NSCLC (non-small cell lung cancer), but may cause acute liver injury and death. The development of highly specific inhibitors for HER4 may prove beneficial in oncology.
Description: Recombinant human HER4 (human epidermal growth factor receptor 4), also known as ErbB4, encompassing amino acids 26-651 (extracellular domain). This construct contains an Fc domain of human IgG1 fused to the C-terminus, followed by a C-terminal Avi-Tag™. The recombinant protein was enzymatically biotinylated using the Avi-Tag™ and 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_005235.3
Host Cell Line: HEK293
Purity: ≥85%
Storage Stability: At least 6 months at –80°C.
Tags: Fc-Avi
Target: HER4
Uniprot: Q15303
Unit Definition: Protein
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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