ArtNr |
PEL-ErbB2-Y1112-2 |
Hersteller |
Raybiotech
|
Menge |
2 Plate Kit |
Kategorie |
|
Typ |
Elisa |
Specific against |
other |
ECLASS 10.1 |
32160605 |
ECLASS 11.0 |
32160605 |
UNSPSC |
41116126 |
Lieferbar |
|
Specificity |
The antibody pair provided in this kit recognizes Human ErbB2 phosphorylated at site Tyrosine-1115. |
Species Detected |
Human |
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 |
ERBB2HER2MLN19NEUNGL |
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 H R P T M B One- Step Substrate |
Other Materials Required |
Distilled or deionized water100 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 R T or O/ N at 4 ° C. Add 100 µl of prepared primary antibody to each well. Incubate 1 h at R T. Add 100 µl of prepared 1 X H R P- Streptavidin to each well. Incubate 1 h at R T. Add 100 µl of T M B One- Step Substrate Reagent to each well. Incubate 30 min at R T. Add 50 µl of Stop Solution to each well. Read at 450 nm immediately. |
Product Features |
Rapidly measure phosphorylated protein in lysates Screen numerous different cell lysates without performing a Western Blot analysis Minimal hands-on time, convenient, and non-radioactive material |
Protein Name & Synonyms |
Receptor tyrosine-protein kinase erbB-2 (EC 2.7.10.1) (Metastatic lymph node gene 19 protein) (MLN 19) (Proto-oncogene Neu) (Proto-oncogene c-ErbB-2) (Tyrosine kinase-type cell surface receptor HER2) (p185erbB2) (CD antigen CD340) |
Pathway |
HER/ErbB SignalingTyrosine Kinase Family |
Introduction |
RayBio® Phospho-ErbB2 (Y1112) ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated ErbB2 protein 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 Blotting analysis.This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-ErbB2. An anti-pan ErbB2 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and ErbB2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-ErbB2 (Y1112) antibody is used to detect phosphorylated ErbB2. 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 ErbB2 (Y1112) 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 20 min. Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI. Reagent Preparation for details. |
Recombinant Human EGF Stimulation of A431 Cell Lines |
A431 cells were untreated or treated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed using this phosphoELISA and Western Blot. |
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