Thermo Fisher Scientific Phospho-NFkB p65 (Ser529) Monoclonal Antibody (B33B4WP), PerCP-eFluor 710, eBioscience

Applications and Tested Dilution

View 4 publications

Product Specifications

Additional Formats:

• eFluor 450
• eFluor 660
• PE
• PE-Cyanine7
• Request custom conjugation

Product Specific Information

The B33B4WP monoclonal antibody recognizes human NFκB p65 subunit phosphorylated at serine 529. NFκB is a transcription factor regulating genes involved in proliferation, apoptosis, immunity, and cancer. Functional NFκB forms homo- or heterodimers from five family members (p65, c-Rel, RelB, p50, p52). Its activity is inhibited by IκB binding, which sequesters NFκB in the cytoplasm. Upon stimulation (e.g., TNFα, IL-1β, LPS, PMA), IκB is degraded, allowing NFκB nuclear translocation and gene regulation. Phosphorylation of p65 at Ser529 or Ser536 enhances NFκB activity.

The B33B4WP antibody detects a 65 kDa band in TNFα-stimulated HeLa cell lysates.

Applications Reported: Intracellular staining followed by flow cytometric analysis.
Applications Tested: Pre-titrated for intracellular staining of human peripheral blood cells at 5 µL (0.03 µg) per test. One test = antibody amount for 100 µL final volume (10⁵–10⁸ cells/test).

Fluorochrome Properties:
PerCP-eFluor® 710 emits at 710 nm and is excited by the 488 nm blue laser. Recommended filter: 710/50; alternative: 695/40. Ensure instrument compatibility.

Light Sensitivity: Protect from light to prevent photo-oxidation.
Fixation: Samples may be stored up to 3 days at 4°C in IC Fixation Buffer (#00-8222) or 1-step Fix/Lyse Solution (#00-5333).

Recommended Protocols:

• Protocol A: Two-step protocol (cytoplasmic proteins)
• Protocol C: Two-step protocol (Fixation/Methanol)
• Protocol B (one-step, nuclear proteins) not applicable

Refer to Staining Intracellular Antigens for Flow Cytometry Protocol for details.

Excitation: 488 nm
Emission: 710 nm
Laser: Blue
Filtration: 0.2 µm post-manufacturing filtered

Target Information

Nuclear factor κB p65 (NF-κB p65), encoded by the RELA gene on human chromosome 11, is also known as RelA. It forms heterodimers with p50 or p52 and is a key regulator of immune and inflammatory responses. TNFα receptor activation leads to IKK-mediated phosphorylation and degradation of IκB, enabling NF-κB nuclear translocation and transcriptional activation, including immunoglobulin κ gene transcription in mature B cells.

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

제품 이미지