Thermo Fisher Scientific Phospho-S6 (Ser235, Ser236) Monoclonal Antibody (cupk43k), PerCP-eFluor 710, eBioscience

Applications and Tested Dilution

• Application: Flow Cytometry (Flow)
• Tested Dilution: 5 µL (0.125 µg)/test
• Publications: View 2 publications

Product Specifications

Available Formats

• Unconjugated
• APC
• eFluor 450
• PE
• PE-Cyanine7
• 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 at serine 235 (human) and serine 236 (mouse).
Ribosomal protein S6 is part of the 40S ribosomal subunit and becomes phosphorylated upon 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 during growth arrest.

• Specificity: Determined by western blotting
• Applications Reported: Intracellular staining and flow cytometric analysis
• Applications Tested: Pre-titrated for 5 µL (0.125 µg) per test in a 100 µL sample volume (10⁵–10⁸ cells/test)

Recommended Protocols

• Protocol A: Two-step for cytoplasmic proteins (surface & intracellular detection)
• Protocol B: One-step for nuclear proteins (transcription factors)
• Protocol C: Two-step Fixation/Methanol for phospho-specific signaling discrimination
  Detailed protocols available in Staining Intracellular Antigens for Flow Cytometry Protocol (BestProtocols®).

Fluorophore Information

• Excitation: 488 nm
• Emission: 710 nm
• Laser: Blue Laser
• Recommended Filter: 710/50 bandpass (695/40 acceptable)
• Light Sensitivity: Protect from light to avoid photo-induced oxidation

Fixation and Storage

Samples can be stored up to 3 days at 4°C in the dark using:

• IC Fixation Buffer (Product #00-8222)
• 1-step Fix/Lyse Solution (Product #00-5333)
  Clone-specific performance should be verified empirically.
• Filtration:* 0.2 µm post-manufacturing filtered.

Target Information

Ribosomes catalyze protein synthesis and consist of small (40S) and large (60S) subunits. The S6 protein, part of the 40S subunit, belongs to the S6E family and is a major substrate for multiple kinases. Phosphorylation occurs at C-terminal serine residues in response to growth factors and mitogens, while dephosphorylation accompanies growth arrest. The protein may regulate cell growth and proliferation by selective mRNA translation. Multiple processed pseudogenes exist throughout the genome.

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

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