H/M/R Phosph Erk1 (Thr202/Tyr204) + Erk2 (Tyr185/Tyr187) & Total Erk1/2 ELISA Kit 1×96-Well
Antigen Information
Accession Number:
- P27361
- P28482
Gene ID:
- 5595 and 5594
Gene Symbol:
- and
- ERK1
- ERK2
- MAPK1
- MAPK3
- PRKM1
- PRKM2
- PRKM3
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)
Assay Format
Pathway
- MAPK Signaling
Specificity
The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Erk1 (pThr202/pTyr204) + Erk2(Tyr185/187) and pan Erk1/2.
Number of Targets Detected
2
Species Detected:
- Human
- Mouse
- Rat
Compatible Sample Types:
- Cell Lysates
- Tissue Lysates
Design Principle
- Sandwich-based
Method of Detection
Colorimetric
Quantitative/Semi-Quantitative:
Semi-Quantitative
Solid Support:
96-well Microplate
Product Specifications
Size
1, 2, or 5 x 96-Well Strip Microplate Kit
Product Features
- Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
- 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
- Anti-Pan Antibody
- HRP-Conjugated Secondary Antibody
- Streptavidin-Conjugated HRP
- 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.
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.
Usage
This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.
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