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蚂蚁淘在线 / 品牌中心 / RayBiotech / RayBiotech/Human, Mouse and Rat Phosphotyrosine MEK1 ELISA/1 Plate Kit/PEL-MEK1-Y-1
商品描述
Antigen Information
Accession Number:
- P31938
- Q01986
- Q02750
Gene ID:
- 5604
Gene Symbol:
- MAP2K1
- MEK1
- PRKMK1
Protein Name & 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)
Assay Format
Pathway:
- Tyrosine Kinase Family
Specificity:
The antibody pair provided in this kit recognizes human tyrosine-phosphorylated MEK1
Number of Targets Detected:
1
Species Detected:
- Human
- Mouse
- Rat
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
Product Specifications
Size:
1, 2, or 5 x 96-Well Strip Microplate Kit
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
Application Notes
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 1X 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.
Typical Data
Positive Control
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 (see Reagent Preparation step 4).
Pervanadate Stimulation of HeLa Cell Line
HeLa cells were untreated or treated with Pervanadate for 10 min at 37°C. Cell lysates were analyzed using this phosphoELISA:
Storage/Stability
Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment.
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Manual
SDS
