Cell-Based H/M/R ERK1/2 (Thr202/Tyr204) JNK (Thr183/Tyr185) p38 MAPK (Thr180/Tyr182) 2×96
Antigen Information
Accession Number:
- P27361
- P28482
- P45983
- Q16539
Gene Symbol:
- CSBP
- CSBP1
- CSBP2
- CSPB1
- ERK1
- ERK2
- JNK1
- MAPK3
- MXI2
- PRKM1
- PRKM2/MAPK8
- PRKM3/MAPK1
- PRKM8
- SAPK1
- SAPK1C/MAPK14
- SAPK2A
Protein Name & Synonyms
Mitogen-activated protein kinase 3 (MAP kinase 3) (MAPK 3) (EC 2.7.11.24) (ERT2) (Extracellular signal-regulated kinase 1) (ERK-1) (Insulin-stimulated MAP2 kinase) (MAP kinase isoform p44) (p44-MAPK) (Microtubule-associated protein 2 kinase) (p44-ERK1)/Mitogen-activated protein kinase 1 (MAP kinase 1) (MAPK 1) (EC 2.7.11.24) (ERT1) (Extracellular signal-regulated kinase 2) (ERK-2) (MAP kinase isoform p42) (p42-MAPK) (Mitogen-activated protein kinase 2) (MAP kinase 2) (MAPK 2)/Mitogen-activated protein kinase 8 (MAP kinase 8) (MAPK 8) (EC 2.7.11.24) (JNK-46) (Stress-activated protein kinase 1c) (SAPK1c) (Stress-activated protein kinase JNK1) (c-Jun N-terminal kinase 1/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 Erk1 phosphorylated at sites Thr202/Tyr204, Erk2 phosphorylated at sites Thr185/Tyr187, JNK phosphorylated at sites Thr183/Tyr185 and p38 phosphorylated at sites Thr180/Tyr182. This kit also recognizes total Erk1/2, total JNK and total p38 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
Two 96-Well Microplate Kit
Product Features
- Site and signal
Pathway
-specific
- In vitro detection of adherent cell culture
- No sample lysis 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
- Cell Culture Microplate
- Wash Buffer A
- Wash Buffer B
- Fixing Solution
- HRP-Conjugated Secondary Antibody
- Blocking Buffer
- TMB One-Step Substrate
- Stop Solution
- Quenching Buffer
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
-20°C
Usage
This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.
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