Cell-Based Human/Mouse/Rat MAPK (Thr180/Tyr182) Phosphorylation ELISA Kit 1 x 96-Well Kit
Antigen Information
Accession Number:
- Q16539
Gene ID:
- 1432
Gene Symbol:
- CSBP
- CSBP1
- CSBP2
- CSPB1
- MAPK14
- MXI2
- SAPK2A
Protein Name & Synonyms
Mitogen-activated protein kinase 14 (MAP kinase 14) (MAPK 14) (EC 2.7.11.24) (Cytokine suppressive anti-inflammatory drug-binding protein) (CSAID-binding protein) (CSBP) (MAP kinase MXI2) (MAX-interacting protein 2) (Mitogen-activated protein kinase p38 alpha) (MAP kinase p38 alpha) (Stress-activated protein kinase 2a) (SAPK2a)
Assay Format
Pathway
- MAPK Signaling
Specificity
The antibodies provided in this kit recognizes human, mouse and rat P38 MAPK phosphorylated at sites Thr180 and pTyr182 as well as total MAPK for comparison.
Number of Targets Detected
1
Species Detected:
- Human
- Mouse
- Rat
Compatible Sample Types:
- Adherent Cell Culture
Design Principle
- Cell-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 Microplate Kit
Product Features
- Site and signal
Pathway
-specific
- In vitro detection of adherent cell culture
- No sample lysis is needed
- Compatible with a standard ELISA plate reader
- Faster results than with ELISA
- Adaptable for high-throughput screening and drug discovery
Application Notes
Kit Components
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
- Seed 10,000-30,000 cells into each well and incubate overnight.
- Apply various treatment, inhibitors or activators according to manufacture’s instructions.
- Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
- Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
- Add 200 µl of Blocking Solution and incubate for 1 h at 37 °C.
- Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
- Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 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.
Storage/Stability
The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.
Usage
This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.
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