Description
Human, Mouse and Rat Phosphotyrosine MEK1 PharmaGenie ELISA Kit (SBRS1890)
Product SKU: | SBRS1890 |
Size: | 96T |
Application: | The antibody pair provided in this kit recognizes human tyrosine-phosphorylated MEK1. |
Uniprot: | P31938 / Q01986 / Q02750 |
Gene ID: | 5604 |
Pathway: | Tyrosine Kinase Family |
Synonyms: | Dual specificity mitogen-activated protein kinase kinase 1 (MAP kinase kinase 1) (MAPKK 1) (MKK1) (EC 2.7.12.2) (ERK activator kinase 1) (MAPK/ERK kinase 1) (MEK 1) |
Target Species: | Human, Mouse, Rat |
Compatible Sample Types: | Cell Lysates, Tissue Lysates |
Design Principle: | Sandwich-based |
Method of Detection: | Colorimetric |
Quantitative/Semi-Quantitative: | Semi-Quantitative |
Storage/Stability: | Upon receipt, the kit should be stored at -20°C. Please use within 6 months from the date of shipment. |
- 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 and equipment required:
The Assay Genie Human, Mouse and Rat Phosphotyrosine MEK1 PharmaGenie ELISA Kit (SBRS1890) will require other equipment and materials to carry out the assay. Please see list below for further details.
- 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 ul 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
- Prepare all reagents and samples as instructed in the manual.
- Add 100 ul of sample or positive control to each well.
- Incubate 2.5 h at RT or O/N at 4 °C.
- Add 100 ul of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 ul of prepared 1X HRP-Streptavidin to each well.
- Incubate 1 h at RT.
- Add 100 ul of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 ul of Stop Solution to each well.
- Read at 450 nm immediately.
HeLa cells were untreated or treated with Pervanadate for 10 min at 37°C. Cell lysates were analyzed using this phosphoELISA: | |
HeLa cells were treated with Pervanadate at 37°C for 10 min. Cells were solubilzed at 4 x 107 cells/ml in lysis buffer. Serial dilutions of lysates were analyzed in this ELISA |