Epidermal Growth Factor, Human (EGF) BioAssay™ ELISA Kit
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The Human Epidermal Growth Factor (Hu EGF) ELISA is to be used for the in vitro quantitative determination of human EGF in human serum, plasma, buffered solution, cell culture medium or urine. The assay will recognize both natural and recombinant human EGF.
Epidermal Growth Factor (EGF) is a 53-amino acid polypeptide originally discovered in the mouse submaxillary gland as an agent that induced precocious eyelid opening and early incisor eruption in the newborn mouse. EGF has been subsequently shown to elicit an array of biological responses that are mediated by specific binding to an EGF receptor/tyrosine kinase located on the cell surface. EGF induces cell proliferation, regulates tissue differentiation, modulates organogenesis, promotes angiogenesis, and accelerates wound healing. It also acts as an inhibitor of gastric acid secretion. EGF is a member of a family of EGF-related growth factors including TGFa, heparin binding EGF-like growth factor (HB-EGF), epiregulin, amphiregulin (AR), betracellulin (BTC), neuregulin 1, neuregulin 2, and neuregulin 3. EGF is initially synthesized as a precursor of a 160-170kD glycoprotein. The mature and soluble form of EGF is a 6kD protein which can be detected in many tissues and nearly all body fluids. High levels of EGF exist in some body fluids such as the saliva, urine, gastric juice, and mammary and seminal fluids. EGF, triggers an intracellular signaling cascade through binding to EGF receptor (also known as ErbB1), causing changes within the target cells.
The EGF receptor, a receptor tyrosine kinase, upon stimulation by EGF results in the phosphorylation of its distinct tyrosine residues and phosphorylates the adaptor protein, SHC. Subsequently, the complex activates the Ras/mitogen activated protein (MAP) kinase pathway.
This cascade couples EGF stimulation to gene transcription.
Serum, Plasma, Urine, Supernatants
The Invitrogen Human Epidermal Growth Factor (Hu EGF) ELISA is to be used for the in vitro quantitative determination of Hu EGF in human serum, plasma, buffered solution, cell culture medium or urine. The assay will recognize both natural and recombinant Hu EGF.
4 hours at RT
Buffered solutions of a panel of substances at 10,000 pg/ml were assayed with the Invitrogen human EGF kit. The following substances were tested and found to have no cross-reactivity: human IL-1b, IL-2, IL-4, IL-6, IL-10, FGF acidic, FGF basic, G-CSF, GM-CSF, IFN-y, KGF, PDGF-AB, SCF, TGF-a, TGF-b1, TNF-a, TNF-b, VEGF-121, VEGF-165; mouse IL-1b, IL-6, G-CSF, GM-CSF, IFN- , TNF-a; rat IL-1a, IL-6, GM-CSF, IFN-y, TNF-a.
The human EGF kit is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). A polyclonal antibody specific for human EGF has been coated onto the wells of the microtiter
strips provided. Samples, including standards of known human EGF content, control specimens, and unknowns, are pipetted into these wells. During the first incubation, the Hu EGF antigen binds to the immobilized (capture) antibody on one site. After washing, a biotinylated monoclonal antibody specific for Hu EGF is added. During the second incubation, this antibody binds to the immobilized Hu EGF captured during the first incubation. After removal of excess second antibody, Streptavidin-Peroxidase enzyme) is added. This binds to the biotinylated antibody to complete the four-member sandwich. After a third incubation and washing to
remove all the unbound enzyme, a substrate solution is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of Hu EGF present in the original specimen.
Inter-assay CV: 5.9%
Intra-assay CV: 5.0%
*E3374-15H1: Hu EGF Standard, purified recombinant Hu EGF expressed in E. coli. Refer to vial label for quantity and reconstitution volume. 2vials, 4 vials
E3374-15H2: Standard Diluent Buffer 15mM sodium azide 1x25ml, 2x25ml
E3374-15H3: Plate Covers, adhesive strips.
E3374-15H4: Hu EGF Antibody-Coated Wells, 96 wells per plate. 1plate, 2plates
Hu EGF Biotin Conjugate (Biotin-labeled anti-EGF). Contains 15 mM sodium azide; 1x11ml, 2x11ml
E3374-15H5: Streptavidin-Peroxidase (HRP), (100x) concentrate. Contains 3.3 mM thymol; 1x0.125ml, 2x125ml
E3374-15H6: Streptavidin-Peroxidase (HRP) Diluent. Contains 3.3mM thymol; 1x25ml
E3374-15H7: Wash Buffer Concentrate (25x); 1x100ml
E3374-15H8: Stabilized Chromogen, Tetramethylbenzidine (TMB) 1x25ml
E3374-15H9: Stop Solution 1x25ml
Storage and Stability:
Store all components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.