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Human Phosphotyrosine TIE2 ELISA

Hersteller Raybiotech
Kategorie
Typ Elisa-Kit
Specific against Human
Menge 2 Plate Kit
Host Hamster - Armenian
ArtNr PEL-TIE2-Y-2
Targets TEK
eClass 6.1 32160605
eClass 9.0 32160605
Lieferbar
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
TEKTIE2VMCMVMCM1
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.
Protein Name & Synonyms
Angiopoietin-1 receptor (EC 2.7.10.1) (Endothelial tyrosine kinase) (Tunica interna endothelial cell kinase) (Tyrosine kinase with Ig and EGF homology domains-2) (Tyrosine-protein kinase receptor TEK) (Tyrosine-protein kinase receptor TIE-2) (hTIE2) (p140 TEK) (CD antigen CD202b)
Pathway
AngiogenesisTyrosine Kinase Family
Introduction
RayBio® Phospho-TIE-2 ELISA 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 phospho-TIE-2 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 phospho-TIE-2. An anti-TIE-2 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated and unphosphorylated TIE-2 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-TIE-2 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Positive Control
K562 cells were treated with Pervanadate 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.
Pervanadate Stimulation of HUVEC Cell Line
HUVEC cells were treated or untreated with Pervanadate for 10 min at 37°C. Cell lysates were analyzed using this phosphoELISA

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Alle Produkte sind nur für Forschungszwecke bestimmt. Nicht für den menschlichen, tierärztlichen oder therapeutischen Gebrauch.

Menge: 2 Plate Kit
Lieferbar: In stock
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Lieferung vsl. bis 30.05.2024 

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