EGFR Antikörper

Produktnummer ABIN98862

Produktdetails anti-EGFR Antikörper

Highlights:

• High quality EGFR Primärantikörper for the detection of EGFR.
• Reliable product with high standard validation data

Target: EGFR (Epidermal Growth Factor Receptor (EGFR))

Reaktivität: Human

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser EGFR Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), ELISA, Immunoprecipitation (IP)

Produktmerkmale: Concentration Definition: by Refractometry

Immunogen: This whole rabbit serum was prepared by repeated immunizations with a peptide synthesized using conventional technology. The sequence of the epitope maps to a region near the carboxy terminus which is identical in human, mouse and rat EGFR.

Produktspezifische Information:

What can the EGFR Antibody ABIN98862 be used for?: This polyclonal EGFR Antibody detects EGFR. The EGFR Antibody has been validated for various applications and can be used for the detection of EGFR and derivatives by Western Blotting, ELISA, Immunoprecipitation.

What validation data is available for this EGFR Antibody?: The Primärantikörper is referenced in 3 publications and is characterized by a proven, very high reliability. It has currently 2 product images that show its performance in a variety of applications. The product is currently available in 250 μL quantity. EGFR Antibody for the detection of EGFR and derivatives.

What is the function of EGFR?: The protein encoded by this gene is a transmembrane glycoprotein that is a member of the protein kinase superfamily. This protein is a receptor for members of the epidermal growth factor family. EGFR is a cell surface protein that binds to epidermal growth factor. Binding of the protein to a ligand induces receptor dimerization and tyrosine autophosphorylation and leads to cell proliferation. Mutations in this gene are associated with lung cancer. Multiple alternatively spliced transcript variants that encode different protein isoforms have been found for this gene. [provided by RefSeq, Jul 2010].

Anwendungsinformationen

Applikationshinweise: Anti-EGFR antibody is specifically designed for ELISA, immunoblotting and immunoprecipitation.  Reactivity in other assays is likely, but has not been determined.  Recognition of EGFR is independent of the phosphorylation status at tyrosine 1173.  No reaction is observed against ErbB-2, ErbB-3 or ErbB-4.   A431 cells, keratinocytes in normal epidermis, or placenta are typically used as positive control sources.  The antigen is typically localized in the cell membrane.   For western blotting, good results are also achieved on PVDF membranes blocked with 5% lowfat milk diluted in TTBS for 1 hour at room temperature.  Also, dilute the primary antibody and secondary in 5% lowfat milk in TTBS.   Anti-EGFR can be diluted up to 1:10,000 for immunoblot depending on the cell line and the amount of EGFR in a particular lysate.  For immunoprecipitation, use approximately 10 µl of the antibody.  The immunoprecipitation mix should contain the antibody, 25 µl of Protein A-agarose beads and 1.0 ml of lysate (lysate contains approximately 1.0 mg of total protein).  This mixture should be rotated overnight at 4°C and then washed 3 times with lysis buffer (used to prepare the lysate).  The resulting bead complex is dissolved in 20-30 µl of 3X SDS-PAGE sample buffer and approximately 15 µl is loaded per lane on an 8% polyacrylamide gel.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #029817 (Western Blotting)

by: ADS Biosystems Inc

No.: #029817

Datum: 18.09.2014

Chargennummer: 19538

Validierte Anwendung: Western Blotting

Positivkontrolle: A549 cells

Negativkontrolle: MCF-7 cells

Bewertung: A strong specific band was observed in the positive control at the expected size (~175 kDa) that is not observed in the negative control.

Validierungsbilder

Figure 1: Scanned image of EGFR (Green) and loading control alpha-tubulin (Red) Western blot using LI-COR Odyssey Infrared Technology. First lane, protein molecular weight markers. Second lane, MCF-7 negative control lysate. Third lane, A549 positive control lysate.

Protokoll

Primärantikörper:

• Antigen: Epidermal Growth Factor Receptor (EGFR)
• Catalog number: ABIN98862
• Lot number: 19538
• Dilution: 1:1,000

Sekundärantikörper:

• Antibody: IRDye 680LT Goat Anti-Rabbit
• Lot number: C30725-01
• Dilution: 1:10,000

Full Protocol:

• Lysates were mixed with NuPAGE® LDS Sample Buffer (Life Technologies NP0007) and NuPAGE® Sample Reducing Agent (Life Technologies NP0004) and denatured for 5 minutes at 90ºC.
• 40 μg of each lysate was electrophoresed on a Bolt 4-12% Bis-Tris Gel (Life Technologies BG04120BOX) and run in Bolt MOPS SDS Running Buffer (Life Technologies B0001) at 160 volts for 1 hour.
• Odyssey Western Protein Standard (LI-COR #928-40000) was run as a molecular weight standard.
• PVDF membrane was activated with methanol.
• Protein samples were transferred to activated PVDF membrane in a wet Bolt Transfer Apparatus (Life Technologies B1000) at room temperature for 1 hour at 20 volts (started at 230mA, ended at 110mA).
• The membrane was blocked in x LI-COR Odyssey WB block solution for 1 hour at room temperature.
• The membrane was incubated with the primary antibody diluted 1:1000 in x LI-COR Odyssey WB block solution incubated 2 hours at room temperature.
• The membrane was washed 4 x 5 minutes in 1 x PBS-T (PBS solution with 0.1% Tween 20).
• The membrane was incubated with IRDye® 800CW Goat anti-Mouse Secondary Antibody (Red) and IRDye 680LT Goat Anti-Rabbit Secondary Antibody (Green) from LI-COR (#827-11081, Lot #C30725-01), both 1:10,000 dilutions. Incubation was performed at room temperature for 45 minutes.
• The membrane was washed 4 x 5 minutes in 1 x PBS-T (PBS solution with 0.1% Tween 20).
• Proteins were detected using Odyssey machine scanning with green channel for loading control and red channel for potential LPL band.

Anmerkungen: - No experimental challenges noted.

Handhabung

Format: Liquid

Konzentration: 85 mg/mL

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Lagerung: -20 °C

Referenzen für anti-EGFR Antikörper (ABIN98862)

Andersson, Anttonen, Färkkilä, Pihlajoki, Bützow, Unkila-Kallio, Heikinheimo: "Sensitivity of human granulosa cell tumor cells to epidermal growth factor receptor inhibition." in: Journal of molecular endocrinology, Vol. 52, Issue 2, pp. 223-34, (2014) (PubMed).

Khoh-Reiter, Jessen: "Evaluation of the cytotoxic effects of ophthalmic solutions containing benzalkonium chloride on corneal epithelium using an organotypic 3-D model." in: BMC ophthalmology, Vol. 9, pp. 5, (2009) (PubMed).

Miettinen, Ustinov, Ormio, Gao, Palgi, Hakonen, Juntti-Berggren, Berggren, Otonkoski: "Downregulation of EGF receptor signaling in pancreatic islets causes diabetes due to impaired postnatal beta-cell growth." in: Diabetes, Vol. 55, Issue 12, pp. 3299-308, (2007) (PubMed).

Details zu EGFR

Target: EGFR (Epidermal Growth Factor Receptor (EGFR))

Andere Bezeichnung: EGFR (EGFR Produkte)

Synonyme: C-erb antikoerper, CG10079 antikoerper, D-EGFR antikoerper, D-Egf antikoerper, DEGFR antikoerper, DER antikoerper, DER flb antikoerper, DER/EGFR antikoerper, DER/faint little ball antikoerper, DER/top antikoerper, DER/torpedo antikoerper, DER1 antikoerper, DEgfr antikoerper, Degfr antikoerper, Der antikoerper, DmHD-33 antikoerper, Dmel\\CG10079 antikoerper, EFG-R antikoerper, EGF-R antikoerper, EGFR antikoerper, EGFr antikoerper, EGfr antikoerper, EK2-6 antikoerper, Egf antikoerper, Egf-r antikoerper, EgfR antikoerper, El antikoerper, Elp antikoerper, Elp-1 antikoerper, Elp-B1 antikoerper, Elp-B1RB1 antikoerper, HD-33 antikoerper, TOP antikoerper, Torpedo/DER antikoerper, Torpedo/Egfr antikoerper, c-erbB antikoerper, d-egf-r antikoerper, dEGFR antikoerper, dEGFR1 antikoerper, dEgfr antikoerper, der antikoerper, egfr antikoerper, flb antikoerper, l(2)05351 antikoerper, l(2)09261 antikoerper, l(2)57DEFa antikoerper, l(2)57EFa antikoerper, l(2)57Ea antikoerper, mor1 antikoerper, top antikoerper, top/DER antikoerper, top/flb antikoerper, torpedo/Egfr antikoerper, torpedo/egfr antikoerper, EGFR12 antikoerper, EGFR15 antikoerper, egfr1 antikoerper, Erbb2 antikoerper, ERBB antikoerper, ERBB1 antikoerper, HER1 antikoerper, PIG61 antikoerper, mENA antikoerper, ErbB-1 antikoerper, Errp antikoerper, 9030024J15Rik antikoerper, AI552599 antikoerper, Erbb antikoerper, Errb1 antikoerper, Wa5 antikoerper, wa-2 antikoerper, wa2 antikoerper, epidermal growth factor receptor antikoerper, Epidermal growth factor receptor antikoerper, epidermal growth factor receptor a (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian) antikoerper, EGFR antikoerper, Egfr antikoerper, egfra antikoerper, egfr1 antikoerper, LOC5564544 antikoerper

Hintergrund: EGFR is a transmembrane glycoprotein that is a member of a family of protein tyrosine kinases crucial to maintaining a normal balance in cell growth and development.  Growth factor receptors are involved not only in promoting the proliferation of normal cells but also in the aberrant growth of many types of human tumors.  For example, the epidermal growth factor receptor (EGFR) is mutated and/or over-expressed in many common solid human squamous cell carcinomas including breast, brain, bladder, lung, gastric, head & neck, esophagus, cervix, vulva, ovary, and endometrium. Over-expression of the EGFR gene occurs in carcinomas with and without gene amplification. EGFR and ErbB-2 are particularly important in breast cancer because increased production or activation has been associated with poor prognosis. EGFR belongs to a family of growth factor receptors, which also includes ErbB-2/HER-2/neu, ErbB-3/HER-3/neu and ErbB-4/HER-4/neu. EGFR can heterodimerize with each of the members of this family.
Synonyms: Receptor tyrosine-protein kinase erbB-1 antibody, c-erbB-1 antibody

Gen-ID: 1956, 29725609

UniProt: P00533

Pathways: NF-kappaB Signalweg, RTK Signalweg, Fc-epsilon Rezeptor Signalübertragung, EGFR Signaling Pathway, Neurotrophin Signalübertragung, Stem Cell Maintenance, Hepatitis C, Positive Regulation of Response to DNA Damage Stimulus, Interaction of EGFR with phospholipase C-gamma, Thromboxane A2 Receptor Signaling, EGFR Downregulation, S100 Proteine