Typical Testing Data/Standard Curve (for reference only)
Epidermal Growth Factor ELISA Kit | EGF elisa kit
Epidermal Growth Factor (EGF)
Typical Testing Data/Standard Curve (for reference only)
Typical Testing Data/Standard Curve (for reference only)
Principle of the Assay: This EGF enzyme-linked immunosorbent assay (ELISA) applies a technique called a quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a monoclonal specific for EGF. Standards or samples are then added to the appropriate microtiter plate wells and incubated. EGF, if present, will bind and become immobilized by the antibody pre-coated on the wells. The microtiter plate wells are thoroughly washed to remove unbound EGF and other components of sample. In order to quantitate the amount of EGF present in the sample, a standardized preparation of horseradish peroxidase (HRP)-conjugated monoclonal antibody specific for EGF is added to each well to "sandwich" the EGF immobilized during the first incubation. The microtiter plate then undergoes a second incubation. The wells are thoroughly washed to remove all unbound HRP-conjugated antibodies and a TMB (3,3'5,5' tetramethyl-benzidine) substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period. Only those wells that contain EGF and enzyme-conjugated antibody will exhibit a change in colour. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the colour change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. In order to measure the concentration of EGF in the samples, this kit contains two calibration diluents (Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant/ urine testing). According to the testing system, the provided standard is diluted (2-fold) with the appropriate Calibrator Diluent and assayed at the same time as the samples. This allows the operator to produce a standard curve of Optical Density (O.D.) versus EGF concentration (pg/mL). The concentration of EGF in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Background/Introduction: Epidermal growth factor (EGF), a polypeptide mitogen, was first observed in 1959 by Cohen and Levi-Montalcini while studying Nerve Growth Factor (NGF) in snake venom extracts.1 It was subsequently isolated and purified from mouse submandibular glands. When injected into new-born mice this new factor caused precocious eyelid opening and incisor eruption.2,3 EGF was further purified, based on its ability to induce the proliferation of basal skin cells.4 Also, a potent inhibitor of gastric acid secretion was identified and isolated from the urine of a pregnant women, and named human beta-urogastrone. It was shown that this protein was very similar to purified human EGF.5,6 The cDNA cloning of mature EGF revealed that this polypeptide (6 kDa, 53 amino acids) belonged to a family of growth factors (TGF-alpha, vaccinia virus growth factor and amphiregulin) that bind to the same 160-185 kDa family of cell surface receptors. Structurally EGF is homologous to TGF-alpha.7,8,9 Mature EGF is formed when the large precursor molecule (pre-EGF), a transmembrane protein (130 kDa, 1217 amino acids) which contains at least seven EGF-like sequences is cleaved. Within this precursor molecule, it is the sequence closest to the C-terminus of the mRNA that represents the coding region for the mature, soluble EGF sequence.7,10 The uncleaved precursor shows biological activity and is capable of binding to the EGF receptor.11,12 The physiological significance of the EGF precursor, nor the mechanisms of its processing into mature EGF have yet to be illustrated. However it is a source for soluble EGF and may be involved in cell to cell signalling or "juxacrine" action by negotiating intercellular communication between membrane-anchored EGF and cells with EGF receptors on their surfaces.7,11 A soluble form of the EGF precursor molecule has been shown to be a heparin-binding protein, upon cleavage and release of the precursor extracellular domain.12 EGF is found in a variety of bodily fluids and tissues, but lower amounts are generally found in the latter. Substantial quantities of EGF are seen in saliva, bile, pancreatic juice, amniotic fluid, mammary fluids and secretions, gastric and duodenal contents, prostatic and seminal fluids, and urine.7,11 Lower amounts of EGF can be found in whole blood and platelet poor plasma, when compared to other body fluids. EGF may be released from platelets during blood coagulation, since EGF levels are increased in serum, when compared to plasma.13 EGF stimulates the growth of many tissues3 and contributes to a wide variety of in vitro and in vivo biological effects, which are varied and depend on the target tissue. In vivo, EGF promotes angiogenesis, liver regeneration, epithelial development, acceleration of wound healing, and inhibits gastric acid secretion.9 It has been shown that EGF in vitro promotes colony formation of epithelial cells in culture,14 suppress insulin-induced glucokinase (hexokinase IV) activity,15 promotes chemomigration of prostate tumor cell line(16) and is a mitogen for fibroblasts and endothelial cells.17-18 Thus, EGF may play a role in human health and disease and could have potential therapeutic role in wound healing, regeneration of liver tissue and repair, embryonic and infant development, and prevention of vaccinia virus infection.9
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a member of the epidermal growth factor superfamily. The encoded protein is synthesized as a large precursor molecule that is proteolytically cleaved to generate the 53-amino acid epidermal growth factor peptide. This protein acts a potent mitogenic factor that plays an important role in the growth, proliferation and differentiation of numerous cell types. This protein acts by binding the high affinity cell surface receptor, epidermal growth factor receptor. Defects in this gene are the cause of hypomagnesemia type 4. Dysregulation of this gene has been associated with the growth and progression of certain cancers. Alternate splicing results in multiple transcript variants.[provided by RefSeq, May 2010]
Uniprot Description
EGF: EGF stimulates the growth of various epidermal and epithelial tissues in vivo and in vitro and of some fibroblasts in cell culture. Magnesiotropic hormone that stimulates magnesium reabsorption in the renal distal convoluted tubule via engagement of EGFR and activation of the magnesium channel TRPM6. Defects in EGF are the cause of hypomagnesemia type 4 (HOMG4); also known as renal hypomagnesemia normocalciuric. HOMG4 is a disorder characterized by massive renal hypomagnesemia and normal levels of serum calcium and calcium excretion. Clinical features include seizures, mild-to mederate psychomotor retardation, and brisk tendon reflexes. 2 isoforms of the human protein are produced by alternative splicing.
Protein type: Hormone; Membrane protein, integral; Ligand, receptor tyrosine kinase
Chromosomal Location of Human Ortholog: 4q25
Cellular Component: extracellular space; lysosomal membrane; integral to membrane; plasma membrane; extracellular region
Molecular Function: protein binding; transmembrane receptor protein tyrosine kinase activator activity; growth factor activity; calcium ion binding; epidermal growth factor receptor binding
Biological Process: negative regulation of epidermal growth factor receptor signaling pathway; peptidyl-tyrosine phosphorylation; transmembrane receptor protein tyrosine kinase activation (dimerization); nerve growth factor receptor signaling pathway; activation of MAPKK activity; positive regulation of transcription, DNA-dependent; signal transduction; positive regulation of MAP kinase activity; platelet degranulation; positive regulation of epidermal growth factor receptor activity; positive regulation of cell proliferation; angiogenesis; DNA replication; STAT protein nuclear translocation; positive regulation of DNA binding; positive regulation of granule cell precursor proliferation; epidermal growth factor receptor signaling pathway; platelet activation; fibroblast growth factor receptor signaling pathway; phosphoinositide-mediated signaling; positive regulation of mitosis; branching morphogenesis of a tube; innate immune response; negative regulation of secretion; blood coagulation; positive regulation of phosphorylation
Disease: Hypomagnesemia 4, Renal
Research Articles on EGF
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Product Notes
The EGF egf (Catalog #AAA590002) is an ELISA Kit and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "Epidermal Growth Factor, ELISA Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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