ArtNr |
PEL-EPHA5-Y-2 |
Hersteller |
Raybiotech
|
Menge |
2 Plate Kit |
Quantity options |
1 Plate Kit
2 Plate Kit
5 Plate Kit
|
Kategorie |
|
Typ |
Elisa-Kit |
Specific against |
Human |
Host |
Hamster - Armenian |
ECLASS 10.1 |
32160605 |
ECLASS 11.0 |
32160605 |
UNSPSC |
41116126 |
Similar products |
EphA5 |
Lieferbar |
|
Specificity |
The antibody pair provided in this kit recognizes Human Tyrosine-Phosphorylated-EphA5. |
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 |
BSKEHK1EPHA5HEK7TYRO4 |
Kit Components |
Pre- Coated 96-well Strip Microplateli id=""> 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 |
Ephrin type-A receptor 5 (EC 2.7.10.1) (Brain-specific kinase) (EPH homology kinase 1) (EHK-1) (EPH-like kinase 7) (EK7) (hEK7) |
Introduction |
RayBio® Phospho-EphA5 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-EphA5 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-EphA5. An anti-EphA5 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated and unphosphorylated EphA5 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-EphA5 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. |
Positive Control |
Jurkat 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 U251 Cell Line |
U251 cells were treated or untreated with Pervanadate for 10 min at 37°C. Cell lysates were analyzed using this phosphoELISA |
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