rev:        January 31, 2005

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ANTIBODIES  

(anti-Human and others as indicated)

RDI Divison of Fitzgerald Industries Intl  offers a wide line of  antibodies. Since no one antibody works best for all applications (neutralization, blotting, histochemistry, ELISA, etc), we offer many different types of antibodies to help solve this problem. Please inquire for other applications or types of antibodies not listed below.


Anti-EGFR  Antibodies (polyclonals)

(see also monoclonals)


Rabbit anti-EGFR (Human/Mouse/Rat)

cat# RDI-EGFRCabrX     $500.00

Presentation: 0.25ml sterile filtered neat sera

Immunogen: synthetic peptide aa 1168-1181. NH2-C-S-L-D-N-P-D-Y-Q-Q-D-F-F-P-K-E-COOH This maps to a region near the carboxy-terminus which is identical in human, mouse and rat EGFR. The amino terminal cysteine was ynthesized to facilitate carrier coupling.

Use: -immunoblotting

       -immunoprecipitation:

       -not tested for other applications

Notes: 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.

Specificity: This antibody can detect EGFR from human, mouse and rat sources. Reactivity of this antibody with EGFR from other species is unknown.

Storage: Store vial at -20° C prior to opening. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.

Recommended  

Dilutions :    

This product was assayed by immunoblot using HRP Goat-anti-Rabbit IgG and ECL for detection. A working dilution of 1:2,000 to 1:3,000 is suggested for this assay to detect 170kDa intact EGFR. Good results are achieved when lysates are transferred to a PVDF membrane after SDS-PAGE electrophoresis and are then blocked with 5% lowfat milk diluted in TTBS for 1 hour at room temperature. After generous washing of the membrane with TTBS, the anti-EGRF antibody is added at a 1:2,000 dilution in 5% lowfat milk and is then incubated overnight at 4° C with gentle rocking. The blot is then reacted with HRP Goat   anti-Rabbit IgG (order code #RDI-6111302 $188.00/2mg)   diluted with 5% lowfat milk diluted in TTBS. Anti-EGRF 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 ul of the antibody. The immunoprecipitation mix contained the antibody, 25 ul of Protein A-agarose beads and 1.0 ml of lysate (lysate contains approximately 1.0 mg of total protein). This mixture is 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 20-30ul of 3X SDS-PAGE sample buffer and approximately 15ul is loaded per lane on an 8% polyacrylamide gel. Researchers should determine optimal titers for other applications.

General  Background: 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 found to be mutated and/or overexpressed in many common solid human squamous cell carcinomas including breast, brain, bladder, lung, gastric, head & neck, esophagus, cervix, vulva, ovary, and endometrium. Overexpression of the EGFR gene occurs in carcinomas with and without gene amplification. EGFR is a transmembrane glycoprotein that is a member of a family of protein tyrosine kinases crucial in maintaining a normal balance in cell growth and development. EGFR and erbB-2 are particularly important in breast cancer because increased production or activation has been associated with poor prognosis. Tumor aggressiveness and patient outcome can be directly correlated to their production levels. Inhibitors of EGFR represent an emerging new class of anti-cancer drugs that target the underlying molecular changes involving oncogenes and tumor suppressor genes, genes that play critical roles in the conversion of normal cells into a cancerous state. EGFR belongs to a family of growth factor receptors which also includes HER-2/neu. It can heterodimerize with each of the members of this family.

For Research Use Only


SHEEP anti-EGF RECEPTOR (MULTI-SPECIES REACTIVE)

HUMAN/RAT/Chicken/Mouse/Dog

Catalog#: RDI-EGFRabS

Price: $594.00/vial

Package Size: 200 micrograms IgG in 0.2ml 0.1M Tris-glycine pH7.4 with 0.05% sodiuma zide

Antigen: recombinant human EGFR (partial cytoplasmic domain of EGFR inclusive of region relevant to exon 15-18).

Species: sheep

Lot: see sheet with shipment

Activity: Reacts withthe EGFR of human, rat , mouse, dog and chicken

Uses:

-western blot: 0.5-2.0microgram/ml detects 175kDa EGFR in 50ug A-431 cell lysate

-immunoprecipitation: 10ug/ml/2 Million A-431 cells. does not inhibit tyrosine kinase activity of precipitated EGF

receptor/antibody complex

-immunocytochemistry: 10ug/ml on 4% PFA fixed cells,

Storage: -20 DEG C long term. AVOID FREQUENT FREEZE THAW CYCLES.

Precautions: For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics.


- NOT RESPONSIBLE FOR ANY PATENT INFRINGEMENTS THROUGH USE OR DERIVATION OF THESE PRODUCTS.



RDI Divison of Fitzgerald Industries Intl

34 Junction Square Drive

Concord MA 01742-3049

USA

phone (978) 371-6446 or (800) 370-2222

fax     (978) 371-2266

EMAIL:antibodies@fitzgerald-fii.com

Ordering terms

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