RayBio® Cell-Based Human/Mouse/Rat ERK1/2 (Thr202/Tyr204) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.
Human/Mouse/Rat Phospho-ERK1/2 (T202/Y204) Cell-Based ELISA Kit
Human/Mouse/Rat Phospho-ERK1/2 (T202/Y204) Cell-Based ELISA Kit
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Product Description
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Species | Human, Mouse, Rat |
| Specificity | The antibodies provided in this kit recognizes human, mouse and rat Erk1 phosphorylated at sites Thr202/Tyr204 and Erk2 phosphorylated at sites Thr185/Tyr187 and total Erk1/2 for comparison. |
| Compatible Sample Types | Adherent Cell Culture |
| Accession Number | P27361|P28482 |
| Gene Id | 5594|5595 |
| Gene Symbols | MAPK1, MAPK3, ERK1, PRKM3, ERK2, PRKM1, PRKM2 |
| 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) |
| Quantitative/Semi-Quantitative | Semi-Quantitative |
| Solid Support | 96-well Microplate |
| Design Principle | Cell-based |
| Method Of Detection | Colorimetric |
| Research Area | Post-Translational Modifications, Phosphorylation, MAPK Signaling |
| Shipping Type | Blue ice |
| Storage | -20°C |
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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
citations
GANT61/BI-847325 combination: a new hope in lung cancer treatment
- Species:
- Human
- Sample Type:
- A549 cell
- Year:
- 2022
- Journal Name:
- Medical Oncology
A novel role of alkaline phosphatase in the ERK1/2 dephosphorylation in renal cell carcinoma cell lines: A new plausible therapeutic target
- Species:
- Human
- Sample Type:
- Cell Lysate (Renal carcinoma cells)
- Year:
- 2014
- Journal Name:
- Biochimie
Palmitoylation of TNF alpha is involved in the regulation of TNF receptor 1 signalling.
- Species:
- Human
- Sample Type:
- Conditioned Media (Hela, 3T3L1, ADAM17-deficient (ADAM17zn/zn) and RAW264.7 cell lines)
- Year:
- 2013
- Journal Name:
- Biochimica et Biophysica Acta Molecular Cell Research
- PubMed ID:
- 23159491
Propranolol induces regression of hemangioma cells through HIF-1-mediated inhibition of VEGF-A
- Species:
- Human
- Sample Type:
- Cell Lysate (Hemangioma endothelial cells treated with propranolol)
- Year:
- 2012
- Journal Name:
- Annals of Surgery
- PubMed ID:
- 22580939
Molecular and cellular characteristics of ABCA3 mutations associated with diffuse parenchymal lung diseases in children
- Species:
- Human
- Sample Type:
- Cell Lysate (Adenocarcinomic basal epithelial cell)
- Year:
- 2012
- Journal Name:
- Human Molecular Genetics
SOCS3 Modulates Interleukin-6R Signaling Preference in Dermal Fibroblasts
- Species:
- Mouse
- Sample Type:
- Cell Lysate (Mouse pups fibroblast cells.)
- Year:
- 2012
- Journal Name:
- Journal of Interferon and Cytokine Research
- PubMed ID:
- 22313262
Expression and shedding of endothelial protein C receptor in prostate cancer cells.
- Species:
- Human
- Sample Type:
- Conditioned Media
- Year:
- 2011
- Journal Name:
- Cancer Cell International
- PubMed ID:
- 21320357
Protective cross talk between activated protein C and TNF signaling in vascular endothelial cells: implication of EPCR, noncanonical NF-B, and ERK1/2 MAP kinases.
- Species:
- Human
- Sample Type:
- Cell Lysate
- Year:
- 2011
- PubMed ID:
- 21228323
Visfatin regulates insulin secretion, insulin receptor signalling and mRNA expression of diabetes-related genes in mouse pancreatic b-cells
- Species:
- Mouse
- Sample Type:
- Cell Lysate (Beta-TC6 cells, BTC)
- Year:
- 2010
- Journal Name:
- Journal of Endocrinology
Serotonin Activates Human Monocytes and Prevents Apoptosis
- Species:
- Human
- Sample Type:
- Cell Lysate (Monocytes)
- Year:
- 2007
- Journal Name:
- Journal of Investigative Dermatology
