Human/Mouse/Rat Phospho-ERK1 (T202/Y204) DuoSet IC ELISA
R&D Systems, part of Bio-Techne | Catalog # DYC1825-2
Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
125-8000 pg/mL
Sample Type
Cell Lysates
Reactivity
Human, Mouse, Rat
Human/Mouse/Rat Phospho-ERK1 (T202/Y204) DuoSet IC ELISA Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Available in 2, 5, and 15- (96-well) plate pack sizes
- Economical alternative to Western blot
Product Summary for Human/Mouse/Rat Phospho-ERK1 (T202/Y204) DuoSet IC ELISA
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure phosphorylated human, mouse, and rat ERK1 (T202 / Y204) in cell lysates. An immobilized capture antibody specific for ERK1 binds both phosphorylated and unphosphorylated ERK1. After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.
Product Specifications
Assay Format
96-well strip plate (sold separately)
Sample Volume Required
Cell lysates (100 µL)
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Scientific Data Images for Human/Mouse/Rat Phospho-ERK1 (T202/Y204) DuoSet IC ELISA
Standard Curve
Specificity of Human/Mouse/Rat Phospho-ERK1 (T2002/Y204) DuoSet IC ELISA is Shown by Western Blot.
Lysates prepared from HeLa human cervical epithelial carcinoma cells treated with 200 nM phorbol 12-myristate 13-acetate (PMA) were incubated in wells coated with Phospho-ERK1 Capture Antibody. Unbound material was removed by washing and bound material was solubilized in SDS gel sample buffer. The same lysate and captured proteins were electrophoresed, transferred to a PVDF membrane and immunoblotted with Phospho-ERK1 Detection Antibody. Only a single band corresponding to phosphorylated ERK1 was detected. While present in lysate at greater amounts than ERK1, phosphorylated ERK2 was not present in the captured material.
Quantification of Phosphorylated ERK1 in PMA-treated Human HeLa Cells.
HeLa human cervical epithelial carcinoma cells were treated with 200 nM PMA for the indicated times. Following cell lysis, ERK1 phosphorylated at T202/Y204 was quantified with this DuoSet IC ELISA. The same lysates were also immunoblotted (inset) with either Anti-phospho-ERK1/ERK2 (p-ERK1/ERK2) (Catalog # AF1575) or anti-ERK1 polyclonal antibodies. The DuoSet IC ELISA results correlate well with the relative amounts of phosphorylated ERK1 detected by Western blot (upper molecular weight band of upper blot corresponds to phosphorylated ERK1). The blot with anti-ERK1 antibody indicates that total levels of ERK1 remained constant during incubations with PMA.The quantification of phosphorylated ERK1 with this DuoSet IC ELISA was also determined in cells pretreated with the MEK1/2 inhibitors U0126 and PD98059, which indirectly inhibit phosphorylation of ERK1 at T202/Y204.Kit Contents for Human/Mouse/Rat Phospho-ERK1 (T202/Y204) DuoSet IC ELISA
- Capture Antibody
- Conjugated Detection Antibody
- Calibrated Immunoassay Standard or Control
- Streptavidin-HRP
Other Reagents Required
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Lysis Buffer*
IC Diluent*
Blocking Buffer*
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent
*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: ERK1
ERK1 and ERK2 (also known as MAPK3 and MAPK1) are 44 and 42 kDa Ser/Thr kinases, respectively. They are part of the Ras-Raf-ERK signal transduction cascade often found downstream of growth factor receptor activation. ERK1 and ERK2 were initially isolated and cloned as kinases activated in response to insulin and NGF. They are expressed in most, if not all, mammalian tissues. Dual threonine and tyrosine phosphorylation activate both ERKs, at Thr202/Tyr204 for human ERK1 and Thr185/Tyr187 for human ERK2.
ERK5, also known as Big Mitogen-activated Protein Kinase 1 (BMK1) and MAPK7, is activated by several mechanisms, including receptor tyrosine kinases, G protein-coupled receptors, and osmotic stress. Like ERK1 and ERK2, ERK5 contains the conserved Thr-Glu-Tyr activation motif in its activation loop. Unlike these ERKs, however, ERK5 contains a unique C-terminal domain that regulates its activation and nuclear translocation.
Long Name
Extracellular Signal-regulated Kinase 1
Alternate Names
MAPK3, P44ERK1, p44mapk, PRKM3
Gene Symbol
MAPK3
Additional ERK1 Products
Product Documents for Human/Mouse/Rat Phospho-ERK1 (T202/Y204) DuoSet IC ELISA
Product Specific Notices for Human/Mouse/Rat Phospho-ERK1 (T202/Y204) DuoSet IC ELISA
For research use only
Loading...
Loading...
Loading...