Technical guide & protocol Tech Support

Product Specifications

Analytical sensitivity

<0.3 U/mL

Assay range

1.6-100 U/mL

Sample type/volume

Cell Lysate
10 µL

Hands-on time

1 hr 20 min

Time-to-result

4 hr

Homogenous (no wash)

No

Interassay CV

6.6%

Intraassay CV

4.9%

Instrument

Colorimetric Microplate Reader

Product size

96 Tests

Contents

Pre-coated 96 well plate
Standard
Standard Dilution Buffer
Detection Antibody
anti-rabbit-HRP
HRP Diluent
Wash Buffer
Chromogen
Stop Solution
Adhesive Plate Covers

Shipping conditions

Wet ice

Storage

2-8°C

Protein name

EGFR

Target site-specificity

Phosphorylated Tyr1173

Protein subtype

Receptor Tyrosine Kinases

Protein family

Growth Factors & Receptors, Kinases & Inhibitors

Species (tested)

Human

Assay kit format

Sandwich ELISA Kit

Detector antibody conjugate

HRP

Label or dye

HRP

About This Kit

The Human EGFR (Phospho) [pY1173] ELISA Kit is a solid-phase sandwich Enzyme-Linked Immunosorbent Assay (ELISA) designed to detect and quantify the level of EGFR (Phospho) [pY1173]) in fresh or frozen human cell lysates. The assay recognizes both natural and recombinant human EGFR(Phospho) [pY1173].

Principle of the method

A monoclonal capture antibody specific for EGFR (Phospho) [pY1173]) has been coated onto the wells of the 96-well plate. During the first incubation, standards of known content and unknown samples are pipetted into the wells and the antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for the target protein is added to the wells and serves as a detection antibody by binding to the immobilized protein captured during the first incubation. After washing, a horseradish peroxidase labeled anti-rabbit IgG is added. This binds to the detection antibody to complete the four member sandwich. After a third incubation and washing to remove all the unbound enzyme, a substrate solution (TMB) 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 target protein present in the original specimen and the optical density can be read on a standard microplate reader.

Rigorous validation

Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Target information

EGFR, epidermal growth factor receptor, is a receptor tyrosine kinases that signals in response to various growth factors. Overexpression has been linked to numerous types of cancer and EGFR is the target of both biological and small molecular therapeutics. EGFR is encoded by the EGFR gene located on chromosome 7 in humans. EGFR belongs to the HER/ERbB family of proteins that includes three other receptor tyrosine kinases, ERbB2, ERbB3, ERbB4. EGFR is a transmembrane receptor and binding of its cognate ligands such as EGF (Epidermal Growth Factor) and TGF alpha (Transforming Growth Factor alpha) to the extracellular domain leads to EGFR dimerization followed by autophosphorylation of the tyrosine residues in the cytoplasmic domain. Overexpression is observed in tumors of the head and neck, brain, bladder, stomach, breast, lung, endometrium, cervix, vulva, ovary, esophagus, stomach and in squamous cell carcinoma.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Gene aliases : EGFR, ERBB, ERBB1, HER1, mENA, NISBD2, PIG61

Gene ID : (Human) 1956

Gene symbol : EGFR

Protein Aliases : avian erythroblastic leukemia viral (v-erb-b) oncogene homolog, cell growth inhibiting protein 40, cell proliferation-inducing protein 61, Epidermal growth factor receptor, erb-b2 receptor tyrosine kinase 1, Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1

UniProt ID (Human) P00533

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