Learn About EGF ELISA Kit

EGF Quantikine ELISA Kit

Human EGF Quantikine ELISA Kit DEG00

ELISA kits find their application in quantitative studies of protein content such as cytokines and chemokines apart from that of intracellular signaling proteins. They are also applied for high throughput screening due to rapid generation of results with consistency and ease of analysis.

EGF is human epidermal growth factor is a protein that is involved in regulating cell growth as it binds to its receptor EGFR. The molecular weight of human EGF protein is 6-kDa and consists of approximately 53 amino acid residues. The EGF ELISA kit is used for quantification of EGF protein in human serum, plasma, buffer solution, cell culture medium or urine.

Principle

The EGF ELISA kit is designed to measure the amount of target bound to the matched antibody pair. The microplate contains pre-coating of targeted antibody in the wells. The solution of sample, standard and control are added into the wells. The protein in the solution binds to the immobilized antibody. Then the second antibody also known as detector antibody attaches to the protein molecule and a sandwich is formed. When the complex is formed a solution containing substrate is added to it that reacts with the enzyme-antibody-target complex and produces a signal to a detectable level. The more the target protein present in the sandwich, the more intense signal is produced.

Mechanism Of Action

The reaction starts when EGF (epidermal growth factor) binds to the EGF Receptor. This binding initiates a series of reactions in response to external stimuli with EGFR forming a hetro-dimeric or homo-dimeric complex with proteins from other family members. The newly formed complex initiates a number of signaling pathways containing (SH2) effector proteins. EGF is widely expressed in kidney, cerebrum, prostrate and salivary glands and acts as a factor causing mutation. It is observed that EGF activates up to 4 downstream signaling cascades which include RAS-RAF_MEK_ERK, PI3 kinase-AKT, PLC gamma-PKC and STAT pathways.

Assay Optimization

With EGF ELISA you don’t need to go through a laborious optimization process. Every step for the ELISA assay is optimized, you need to follow the protocol. The plates are pre-coated with antibodies. The important thing to consider is to follow every step of the protocol especially the washing and incubation time of the plates.

Necessary Precautions

Before you start you should know that ELISA kits contain a minute amount of sodium azide. In normal circumstances sodium azide reacts with lead and copper to form metal azide which is explosive. So when you are about to dispose of the reagents, flush the drains with a large amount of water, so that the azides won’t accumulate in the pipes. Avoid touching your eyes and face or ingestion as toxic substances are present in it. Rinse your eyes thoroughly with water in case of touch. Always store the kit between 2-8oC for long term usage.

Materials Provided With ELISA

In the kit all materials and reagents required to perform ELISA are present. These materials include EGF capture antibody coated 96-well plates, calibrated standard, wash buffer and dilution buffer, detection antibody, secondary detection reagents, chromogen (stabilized), stop solution and plate covers.You will get the detailed protocol with the kit.

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