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Antigen Information
Accession Number:
- P27361
- P28482
- P45983
- Q16539
Gene Symbols:
CSBP | CSBP1 | CSBP2 | CSPB1 | ERK1 | ERK2 | JNK1 | MAPK3 | MXI2 | PRKM1| PRKM2/MAPK8 | PRKM3/MAPK1 | PRKM8 | SAPK1 | SAPK1C/MAPK14 | SAPK2A View more
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
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Arai M., et al. Protective Effects of Fucoidan Against Interleukin-1?-induced Inflammation in SW982 Human Synovial Cells. The Showa University Journal of Medical Sciences Vol. 26 (2014) No. 1 p. 47-56. http://dx.doi.org/10.15369/sujms.26.47
Species: Human
Sample type: Cell Lysate -
Chen HM., Baicalein, an active component of Scutellaria baicalensis Georgi, prevents lysophosphatidylcholine-induced cardiac injury by reducing reactive oxygen species production, calcium overload and apoptosis via MAPK pathways. BMC Complementary and Alternative Medicine 2014, 14:233 doi:10.1186/1472-6882-14-233
Species: Rat
Sample type: Cell Lysate -
Matsui S., et al. alcitonin gene-related peptide regulates mitogen-activated protein kinase pathway to decrease transforming growth factor ?1-induced hepatic plasminogen activator inhibitor-1 mRNA expression in HepG2 cells. Biosci Biotech and Biochem. Volume 78, Issue 5, 2014 DOI: 10.1080/09168451.2014.910096
Species: Human
Sample type: Cell Lysate -
Kim S, Yamamoto K, Nakamura Y, et al. A Possible Mechanism of Cisplatin-Induced Tumor Necrosis Factor (TNF)-? Production in Murine Macrophages. Pharmacology & Pharmacy. 2013;4(2):146-151.
Species: Mouse
Sample type: Cell Lysate -
Bas E., Van De Water T., Gupta C., et al. Efficacy of three drugs for protecting against gentamicin-induced hair cell and hearing losses. Br J Pharmacol. 2012 Jul;166(6):1888-904. doi: 10.1111/j.1476-5381.2012.01890.x.
Species: Rat
Sample type: Tissue Lysate -
Allen I., Wilson J., Schneider M., et al. NLRP12 suppresses colon inflammation and tumorigenesis through the negative regulation of noncanonical NF-?B signaling. Immunity. 2012 May 25;36(5):742-54. doi: 10.1016/j.immuni.2012.03.012.
Species: Mouse
Sample type: Cell Lysate -
Patel, Suraj J., et al. "The inflammatory response to double stranded DNA in endothelial cells is mediated by NF?B and TNF?." PloS one 6.5 (2011): e19910.
Species: Human
Sample type: Conditioned Media -
Gonzalez-Sarrias A., et al. NF-kB-dependent anti-inflammatory activity of urolithins, gut microbiota ellagic acid-derived metabolites, in human colonic fibroblasts. British Journal of Nutrition (2010), 104, 503-512
Species: Human
Sample type: Cell Lysate
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Usage
This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.
