Thermo Fisher Scientific Phospho-S6 (Ser235, Ser236) Monoclonal Antibody (cupk43k), eFluor 450, eBioscience

Applications and Tested Dilution

Product Specifications

Additional Formats:

• Unconjugated
• APC
• PE
• PE-Cyanine7
• PerCP-eFluor 710
• Request custom conjugation

Product Specific Information

This cupk43k monoclonal antibody recognizes human and mouse ribosomal protein S6 (40S ribosomal protein S6, phosphoprotein NP33, RPS6, RS6, S6) when phosphorylated on serine 235 (S235, human) and serine 236 (S236, mouse). Ribosomal protein S6 is a component of the 40S subunit of the ribosome and is phosphorylated following stimulation by growth factors, tumor-promoting agents, or mitogens. Phosphorylation by p70S6K and PKDCD upregulates translation of proteins involved in cell cycle entry, while dephosphorylation occurs upon growth arrest.

The specificity of this antibody was confirmed by western blotting.

Applications Reported:

• Intracellular staining followed by flow cytometric analysis.

Applications Tested:

• Pre-titrated and tested for intracellular staining and flow cytometric analysis of stimulated normal human peripheral blood cells.
• Recommended use: 5 µL (0.125 µg) per test (100 µL final volume, 10⁵–10⁸ cells/test).

Protocols:

• Protocol A: Two-step (cytoplasmic) – flexible detection of surface and intracellular proteins.
• Protocol B: One-step (nuclear) – for transcription factors with surface/phospho-cytoplasmic proteins.
• Protocol C: Two-step (Fixation/Methanol) – optimal for phospho-specific signaling discrimination.

All protocols are available in the Staining Intracellular Antigens for Flow Cytometry Protocol (BestProtocols®).

Fluorochrome Information:
eFluor™ 450 is an alternative to Pacific Blue™, excited at 405 nm and emitting at 445 nm (Violet laser compatible).
Filtered post-manufacturing (0.2 µm).

Target Information

Ribosomes catalyze protein synthesis and consist of a small 40S and a large 60S subunit, composed of 4 RNA species and ~80 distinct proteins. The S6 protein, part of the 40S subunit, belongs to the S6E family and is the major substrate of ribosomal protein kinases. Multiple C-terminal serine residues are phosphorylated in response to growth factors, mitogens, and tumor-promoting agents, and dephosphorylated during growth arrest. This protein may regulate cell growth and proliferation through selective translation of specific mRNA classes. Multiple processed pseudogenes exist in the genome.

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

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