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Phospho-ERK1/2 ELISA Assay Kit (Colorimetric)

Phospho Extracellular signal-Regulated Kinase 1/2 (ERK 1/2) ELISA is a non-isotopic immunoassay for the in vitro quantitation of human phospho ERK 1/2 (T202/Y204-T185/Y187) protein in cell lysates. Also called p42/p44 or Mitogen Activated Protein Kinase 1/2 (MAPK1/MAPK2).

 

Features:

- Colorimetric assay
- HTS compatible format
- High sensitivity
- 4 deg C storage with long shelf life
- Non-radioactive
- 5 hour protocol

 
Performance Summary:

- Sensitivity: 40 cells/well
- Intra-assay C.V.'s: <3% Mid-level
- Inter-assay C.V.'s: <4% Mid-level
Ordering Information:
Cat. No.
X1839K
 
Size
1 Plate
 
For larger quantity pricing, please contact Customer Service.

 
Legend: CHO cells transfected with the human S1P4 receptor for S1P (Edg6) were serum stared for 4 hours and challenged with S1P or LPA. PMA was used as a positive control for Phospho ERK 1/2 production. PHOSPHO ERK 1/2 was detemined to be an excellent surrogate marker for stimulation with S1P compared with traditional methods for GPCR signaling such as intracellular Calcium flux.

Protocol Summary: Not to be used in place of detailed protocol:

Steps
Incubation
1. Equilibrate kit to room temperature -
2. Add samples and standards to wells 2 hours room temp. (or overnight 4°C)
3. Wash 4x -
4. Add detector antibody to all wells 2 hours - room temp.
5. Wash 4x -
6. Add HRP conjugate to all wells 30 minutes - room temp.
7. Wash 4x -
8. Add substrate to all wells 30 minutes - room temp.
9. Add stop solution to all wells -
10. Read plate at 450 nm/550 nm -