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Phosphotyrosine EGFR ELISA Kit

Phosphotyrosine EGFR ELISA Kit

Item no.	PEL-EGFR-Y-5
Manufacturer	Raybiotech
Amount	5 Plate Kit
Quantity options	1 Plate Kit 2 Plate Kit 5 Plate Kit
Category	Oncology Neuroscience Neural Signaling By Cell Type Blastoma Carcinoma Glioma Hematology Lymphoma Myeloma Melanoma Sarcoma Adenocarcinoma ELISA ELISA & Immunoassays ELISA Kits Immunoassay Astrocytoma By Organ Bladder Cancer
Type	Elisa-Kit
Specific against	Human (Homo sapiens), Mouse (Murine, Mus musculus)
Host	Hamster - Armenian
ECLASS 10.1	32160605
ECLASS 11.0	32160605
UNSPSC	41116126
Similar products	EGFR, ErbB1, EGFR (P-Tyr1086), ErbB1 (P-Tyr1086)
Shipping Condition	Cool pack
Available
Specificity	The antibody pair provided in this kit recognizes Human Tyrosine-Phosphorylated-EGFR.
Manufacturer - Category	Assay Kits\|Phosphorylation Assays\|ELISA Kits\|Phosphorylation ELISA
Shipping Temperature	Blue ice
Storage Conditions	-20°C
Compatible Sample Types	Cell Lysates Tissue Lysates
Design Principle	Sandwich-based
Method of Detection	Colorimetric
Quantitative/Semi-Quantitative	Semi-Quantitative
Solid Support	96-well Microplate
Gene Symbols	EGFRERBBERBB1HER1
Kit Components	Pre-Coated 96-well Strip Microplate Wash Buffer Biotinylated Anti-Phosphotyrosine Antibody Stop Solution Assay Diluent(s) Positive Control Sample Lysis Buffer Streptavidin-Conjugated HRP TMB One-Step Substrate
Other Materials Required	Distilled or deionized water 100 ml and 1 liter graduated cylinders Tubes to prepare sample dilutions Protease and Phosphatase inhibitors Precision pipettes to deliver 2 µl to 1 ml volumes Adjustable 1-25 ml pipettes for reagent preparation Benchtop rocker or shaker Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline	Prepare all reagents and samples as instructed in the manual. Add 100 µl of sample or positive control to each well. Incubate 2.5 h at RT or O/N at 4°C. Add 100 µl of prepared primary antibody to each well. Incubate 1 h at RT. Add 100 µl of prepared 1 X HRP-Streptavidin to each well. Incubate 1 h at RT. Add 100 µl of TMB One-Step Substrate Reagent to each well. Incubate 30 min at RT. Add 50 µl of Stop Solution to each well. Read at 450 nm immediately.
Protein Name & Synonyms	Epidermal growth factor receptor (EC 2.7.10.1) (Proto-oncogene c-ErbB-1) (Receptor tyrosine-protein kinase erbB-1)
UNSPSC Code	41116158
Manufacturer - Specificity	The antibody pair provided in this kit recognizes Human Tyrosine-Phosphorylated-EGFR.
Pathway	HER/ErbB SignalingTyrosine Kinase Family
Introduction	RayBio® Phospho-EGFR ELISA (Enzyme-Linked Immuno-sorbent Assay) kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cell lysates. By determining phosphor-EGFR in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blot analysis.This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human and mouse phospho-EGFR. An anti-EGFR antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated and unphosphorylated EGFR present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-phosphotyrosine antibody is used to detect only tyrosine-phosphorylated protein. After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of phospho-EGFR bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Positive Control	A431 cells were treated with recombinant human EGF at 37°C for 10 min. Solubilize cells at 4 x 107 cells/ml in lysis buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI Reagent Preparation for detail.
Stimulation	A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed using this phospho-ELISA.

Note: The presented information and documents (Manual, Product Datasheet, Safety Datasheet and Certificate of Analysis) correspond to our latest update and should serve for orientational purpose only. We do not guarantee the topicality. We would kindly ask you to make a request for specific requirements, if necessary.

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