Biolegend, 285209, Spark Red™ 718 anti-human EGFR (Flexi-Fluor™) Antibody, 50μg

Epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein and member of the protein kinase superfamily that regulates cell growth and differentiation. EGFR binds EGF, TGF-α, amphiregulin, betacellulin, heparin-binding EGF-like growth factor, GP30, and vaccinia virus growth factor - all members of the EGF family. Ligand binding induces EGFR dimerization and autophosphorylation, initiating the MAPK, Akt, and JNK signaling pathways. EGFR is expressed by epithelial and endothelial cells and is frequently expressed by epithelial carcinomas.
50μg
Verified Reactivity: Human
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Non-small cell lung cancer (NSCLC) cell line NCI-H322
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation: The antibody was purified by affinity chromatography and conjugated with Spark Red™ 718 under optimal conditions.
Concentration: 0.2 mg/mL
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application: FC
Recommended Usage: Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. * Spark Red™ 718 has a maximum excitation of 697 nm and a maximum emission of 711 nm.
Excitation Laser: Red Laser (633 nm)
Additional Product Notes: For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.
Application References(PubMed link indicates BioLegend citation): Yamaguchi M, et al. 2009. The 15th Annual Meeting Japan Society of Gene Therapy. p1056. Abstract 92.
RRID: AB_3662445 (BioLegend Cat. No. 285209)
Structure: Member of the protein kinase superfamily; transmembrane glycoprotein; ligand binding induces dimerization and autophosphorylation
Distribution: Epithelial cells and endothelial cells; frequently found in epithelial carcinomas
Function: Controls cell growth and differentiation
Interaction: MAPK, Akt, JNK
Ligand/Receptor: Members of the epidermal growth factor (EGF) family such as EGF, TGF-α, amphiregulin, betacellulin, heparin-binding EGF-like growth factor, GP30 and vaccinia virus growth factor
Cell Type: Endothelial cells, Epithelial cells
Biology Area: Cell Biology, Cell Cycle/DNA Replication, Immunology, Innate Immunity, Neuroscience, Synaptic Biology
Molecular Family: CD Molecules, Growth Factors
Antigen References: 1. da Cunha Santos G, et al. 2011. Annu. Rev. Pathol. 6:49. 2. Gusterson BA and Hunter KD. 2009. Lancet Oncol. 10:522. 3. Mano M and Humblet Y. 2008. Nat. Clin. Pract. Oncol. 5:415. 4. Pao W and Chmielecki J. 2010. Nat. Rev. Cancer 10:760.
Gene ID: NA
UniProt: View information about EGFR on UniProt.org
Clone: AY13
Regulatory Status: RUO
Other Names: Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1, HER1
Isotype: Mouse IgG1, κ
Q: What are Flexi-Fluors?
A: Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer.
Q: How quickly will I receive my order?
A: We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary.
Q: How are Flexi-Fluors different from regular catalog products?
A: Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.
Q: How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?
A: Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL. 

For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.
Q: I can’t find the antibody-dye combination that I need. When will it be available?
A: We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services.
Q: I need help to validate the performance of my Flexi-Fluor. Who should I contact?
A: Please get in touch with Technical Service for assistance.
Q: Can I order more than 50 μg of a Flexi-Fluor?
A: Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.
Q: What is the expiration date of my Flexi-Fluor?
A: Expiration dates can be found on the vial label or by using our CoA lookup tool.
Q: What if my requested antibody-dye combination doesn't perform as expected?
A: Flexi-Fluor™ products have been carefully selected by our scientists for their excellent conjugation properties and broad compatibility. In the rare case that an antibody-dye combination does not perform as expected due to the unique characteristics of a specific pairing, please contact our Technical Service team. We will work with you to identify alternative options to meet your panel requirements.