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.

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	-