Thermo Fisher Scientific Phospho-IkB alpha (Ser32, Ser36) Monoclonal Antibody (RILYB3R), PE, eBioscience

Applications and Tested Dilution

Product Specifications

Additional Formats

• eFluor 660
• Request custom conjugation

Product Specific Information

This RILYB3R monoclonal antibody recognizes human and mouse IkB alpha phosphorylated on serines 32 and 36 (S32/S36).
IkB alpha inhibits classical NF-kappaB signaling by masking the nuclear localization signals of NF-kappaB proteins, keeping them inactive in the cytoplasm. Upon stimulation (e.g., T/B cell receptor engagement, TNFα, IL-1), the IKK complex phosphorylates IkB alpha, leading to its degradation and NF-kappaB activation.

Applications Reported: Intracellular staining followed by flow cytometric analysis.
Applications Tested: Pre-titrated and tested for intracellular staining of normal human peripheral blood cells. Recommended use: 5 µL (0.125 µg) per test in 100 µL final volume. Optimal cell number and titration should be determined empirically (10⁵–10⁸ cells/test).

Staining Protocols:

• Protocol A: Two-step intracellular staining for cytoplasmic proteins.
• Protocol C: Fixation/Methanol method for phospho-specific discrimination between unstimulated and stimulated samples.

Detailed protocols are available in the BestProtocols® Flow Cytometry section.

Emission: 578 nm
Laser Compatibility: Blue, Green, Yellow-Green lasers
Filtration: 0.2 µm post-manufacturing filtered

Target Information

IkB-alpha (40 kDa) inhibits NF-κB activity by masking its nuclear localization sequence, preventing nuclear translocation. Activation of NF-κB occurs via phosphorylation and degradation of IkB-alpha, typically at Ser32 and Ser36, mediated by IKKα/β/γ complex.
Stimuli include TNF-α, IL-1β, PMA, LPS, UV, viral infection, and other stress signals. In humans, the gene is located on chromosome 14q.

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

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