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Application: Useful for studying targeted degradation of human EGFR on the cell surface.
Assay Conditions: This product was validated by flow cytometry analysis for its ability to induce EGFR surface degradation in HCT-116 cells. HCT-116 cells were cultured in a 6-well plate and treated with 50 nM of Anti-RNF43-Anti-EGFR AbTAC (#102492) for 6 hours. Negative control groups included HCT-116 cells treated with Anti-RNF43-Anti-EGFR Bispecific AbTAC in the presence of bafilomycin A1 (a lysosomal inhibitor), Anti-RNF43, Avi-Tag, Biotin-Labeled Antibody (#102549), and DMSO. After incubation, cells were analyzed by flow cytometry. One million cells of each sample were blocked and stained with PE anti-human EGFR Antibody (BioLegend #352904) for 30 minutes on ice, washed three times, and analyzed by flow cytometry
Background: RNF43, or ring finger protein 43, is an E3 ubiquitin ligase that acts as a tumor suppressor to negatively regulate the WNT signaling pathway through promoting the degradation of frizzled WNT receptors. AbTACs target both tumor-associated antigens (TAAs) and RNF43 on the cell surface, promoting the ubiquitination and subsequent degradation of TAAs by RNF43. This mechanism results in the suppression of tumor cell proliferation and may enhance the efficacy of anti-cancer therapies in tumors that overexpress TAAs. This bispecific antibody facilitates EGFR degradation by recruiting RNF43, exploiting its E3 ligase activity to trigger targeted degradation.
Description: This anti-EGFR-anti-RNF43 AbTAC (Antibody-based Targeted Protein Degradation) is a purified, recombinant monoclonal antibody that recognizes human EGFR (epidermal growth factor receptor) and human RNF43 (ring finger protein 43). This antibody has been functionally tested in a flow cytometry assay for its ability to induce targeted degradation of EGFR.
Endotoxin Level: ≤27 EU/mg
Format: Aqueous buffer solution
Formulation: 8 mM phosphate, pH 7.4, 110 mM NaCl, 2.2 mM KCl, and 20% glycerol
Host Cell Line: HEK293
Purity: ≥85%
Storage Stability: At least 12 months at -80°C.
Target: EGFR, RNF43
Uniprot: P00533, Q68DV7
Unit Definition: Antibody
Warnings: Avoid multiple freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)