Thermo Fisher Scientific Phospho-ERK1/2 (Thr202, Tyr204) Monoclonal Antibody (MILAN8R), PE, eBioscience

Applications and Tested Dilution

• Application: Flow Cytometry (Flow)
• Tested Dilution: 5 µL (0.125 µg)/test
• Publications: View 10 publications

Product Specifications

Additional Formats

• Unconjugated
• Alexa Fluor 488
• APC
• PE-eFluor 610
• PerCP-eFluor 710
• Request custom conjugation

Product Specific Information

Description:
The MILAN8R monoclonal antibody recognizes human and mouse extracellular signal-regulated kinases 1 and 2 (ERK1/2, also known as p44/p42 or MAPK3/1) when phosphorylated on T202/Y204. ERK1/2 are serine/threonine kinases involved in cellular processes such as proliferation, differentiation, motility, and survival. Activation occurs via MEK1/2 phosphorylation at T202/Y204 (ERK1) and T185/Y187 (ERK2). Downstream targets include p90RSK and Elk-1. ERK1/2 are negatively regulated by MAPK phosphatases (DUSPs/MKPs) and MEK inhibitors (U0126, PD98059). ERK pathway dysregulation is associated with various cancers.

Specificity of the MILAN8R clone was confirmed by ELISA, flow cytometry, and western blotting.

Applications Reported:

• Intracellular staining followed by flow cytometric analysis.

Applications Tested:

• Pre-titrated and validated for intracellular staining and flow cytometric analysis of normal human peripheral blood cells.
• Recommended usage: 5 µL (0.125 µg) per test (100 µL final volume).
• Applicable cell number: 10⁵–10⁸ cells/test.

Staining Protocol:
Use Protocol C (Two-step: Fixation/Methanol).
Protocol A (cytoplasmic proteins) and Protocol B (nuclear proteins) are not compatible.
Detailed procedures are available in BestProtocols® - Staining Intracellular Antigens for Flow Cytometry.

Fluorescence Characteristics:

• Excitation: 488–561 nm
• Emission: 578 nm
• Compatible Lasers: Blue, Green, Yellow-Green
• Filtration: 0.2 µm post-manufacturing filtered

Target Information

ERK1 and ERK2 are widely expressed kinases regulating meiosis, mitosis, and postmitotic functions. They are activated by growth factors, cytokines, viral infection, GPCR ligands, and transforming agents. Upon receptor tyrosine kinase activation, Ras activates Raf, which phosphorylates MEK1/2, leading to ERK1/2 phosphorylation. Activated ERKs phosphorylate cytoplasmic targets or translocate to the nucleus to activate transcription factors such as c-Fos and Elk-1.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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