RayBio® Human/Mouse/Rat Phospho-ERK1 (T202/Y204) and ERK2 (T185/Y187) ELISA Kit quantitatively measures phosphorylated ERK1 (Thr202/Tyr204) and ERK2 (Thr185/Tyr187) in lysate samples.
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Product Description
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Estimated Lead Time | 1-2 business days |
| Species | Human, Mouse, Rat |
| Specificity | The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Erk1 (pThr202/pTyr204) + Erk2 (pTyr185/187). |
| Compatible Sample Types | Tissue Lysates, Cell Lysates |
| 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 | Sandwich-based |
| Method Of Detection | Colorimetric |
| Research Area | Post-Translational Modifications, Phosphorylation, MAPK Signaling |
| Shipping Type | Blue ice |
| Storage | -20°C |
Introduction
RayBio® Phospho-ERK1 (Thr202/Tyr204) and ERK2 (Thr185/Tyr187) Quantitative ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in Human cell lysates. By determining phosphorylated ERK protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human, Mouse and Rat phospho-ERK1/2. An antipan ERK antibody has been coated onto a 96-well plate. Standard and Samples are pipetted into the wells and ERK present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-phospho-ERK antibody is used to detect phosphorylated ERK. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of ERK1 (Thr202/Tyr204) and ERK2 (Thr185/Tyr187) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human, Mouse and Rat phospho-ERK1/2. An antipan ERK antibody has been coated onto a 96-well plate. Standard and Samples are pipetted into the wells and ERK present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-phospho-ERK antibody is used to detect phosphorylated ERK. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of ERK1 (Thr202/Tyr204) and ERK2 (Thr185/Tyr187) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Product Features
- Rapidly measure phosphorylated protein in lysates
- Screen numerous different cell lysates without performing a Western Blot analysis
- Minimal hands-on time, convenient, and non-radioactive material
Application Notes
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Anti-Phospho Antibody
- HRP-Conjugated Secondary Antibody
- Assay Diluent
- TMB One-Step Substrate
- Stop Solution
- Lysis Buffer
- Positive Control Sample
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
- Prepare all reagents and samples as instructed in the manual.
- Add 100 µl of sample or positive control to each well.
- Incubate 2.5 h at RT or O/N at 4 °C.
- Add 100 µl of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared 1X HRP-Streptavidin to each well.
- Incubate 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.
Typical Data
Standard Curve
PDGFBB Stimulation of NIH3T3 Cell Lines
NIH3T3 cells were untreated or treated with PDGFBB for 10 min. Cell lysates were analyzed using this phosphoELISA kit and western blot.
Storage/Stability
Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), HRP-Streptavidin (Item G), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.