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)
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
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
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