Anti-EGFR Biotin

Overview

Anti-EGFR Biotin is a Mouse monoclonal targeting EGFR, supplied as a Biotin format for FC workflows. It supports measurement of Human target expression in common experimental systems.

Key elements and design rationale

• Clone: 528 — consistent clone identity can support panel reproducibility and cross-study comparisons.
• Isotype: IgG2a, k — informs selection of matched controls and secondary reagents when relevant.
• Conjugate: Biotin — enables direct detection in fluorescence-based assays. Excitation is typically matched to Blue (488nm) lasers in cytometer configurations.
• Host species: Mouse — useful for panel design and control strategy planning.
• Reactivity: Human — interpret staining in the context of species-specific sequence and expression differences.

Key specifications such as clone identity, isotype, and fluorophore conjugation help researchers align panel design, control selection, and instrument configuration with the biological question and sample type.

Biological background

The clone 528, a mouse monoclonal antibody, specifically reacts with an epitope of the ~170 kDa extracellular protein domain of human epidermal growth factor receptor or commonly known as EGFR. Physiologically EGFR is expressed in the skin, gastrointestinal system, kidney, and other normal tissues as well as aberrantly over expresses in epithelial cancer cells of lung, pancreas, colon, breast, and on the head and neck squamous cell. EGFR signaling is activated upon binding one of its ligands including epidermal growth factor (EGF), transforming growth factor α (TGF α), Amphiregulin, and heparin binding-EGF (HB-EGF). Upon activation, EGFR becomes an active homodimer from an inactive monomeric form, resulting in several downstream signal transduction cascades including the MAPK, Akt and JNK pathways, leading to DNA synthesis and cell proliferation closely linked with cancer pathogenesis. The 528 antibody has been reported to block EGF binding to its receptor and inhibits A431 tumor formation in nude mice. Therefore, the anti-EGFR monoclonal antibody based anti-cancer immunotherapy has strong clinical potential against various epithelial solid malignant tumors.

Research relevance and current trends

• High-parameter immunophenotyping: combining EGFR with complementary lineage and activation markers to resolve complex cell states.
• Panel standardization and data comparability: increasing emphasis on consistent reagents, compensation-aware fluorophore choices, and shared gating strategies.
• Integration with single-cell multi-omics: pairing surface marker profiling with transcriptomic or proteomic readouts to connect phenotype to function.

Common research applications

• Flow cytometry: quantify EGFR-positive populations and compare expression distributions across conditions or time points.
• Cell sorting: enrich EGFR-defined subsets for downstream RNA/protein assays or functional readouts.

Changes in measured signal are typically interpreted in the context of cell subset frequency, activation/differentiation state, and sample processing effects rather than as a standalone readout.

Notes for experimental interpretation

• Fluorophore selection: consider brightness, spectral overlap, and instrument configuration; compensation and spillover can affect apparent population boundaries.
• Biology-driven confounders: activation state, differentiation, and isoform/PTM variation can shift epitope accessibility and apparent expression.
• Control concepts: include matched isotype and fluorescence-minus-one (FMO) controls where appropriate, and interpret results alongside biological positive/negative reference samples.

For antibody-based assays, monoclonal versus polyclonal format can influence epitope recognition breadth and signal consistency. Conjugated antibodies support direct detection and can simplify multicolor panel design when paired with appropriate controls and instrument settings.

Customization & Add-ons: Can’t find the antibody you need—or require a custom format for your assay? We can help you source the best match or support custom antibody solutions for diverse research needs, including species and isotype selection, conjugations and labeling (e.g., HRP/AP, biotin, fluorophores), purification grade options (Protein A/G, affinity purified), formulation preferences (buffer selection, carrier-free, glycerol-free), custom concentrations and aliquoting, low-endotoxin options for cell-based work, and application-focused QC/validation support (project dependent). Click Talk to a Scientist to submit a request, email us at support@biohippo.com, or explore our Research Services for additional support—our team will follow up with feasibility details and next steps.