Thermo Fisher Scientific Phospho-NFkB p65 (Ser529) Monoclonal Antibody (B33B4WP), eFluor 660, eBioscience
인산화된 NFkB p65 (Ser529)을 특이적으로 인식하는 단일클론 항체입니다. 인간 시료에 반응하며, eFluor 660 형광표지로 유세포분석(Flow Cytometry)에 최적화되어 있습니다. 세포 내 단백질 염색용으로 사전 적정되어 있으며, 연구용으로만 사용 가능합니다.
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Applications and Tested Dilution
| Application | Tested Dilution |
|---|---|
| Flow Cytometry (Flow) | 5 µL (0.06 µg)/test |
Product Specifications
| 항목 | 내용 |
|---|---|
| Species Reactivity | Human |
| Published Species | Human, Mouse |
| Host / Isotype | Mouse / IgG2a, kappa |
| Recommended Isotype Control | Mouse IgG2a kappa Isotype Control (eBM2a), eFluor™ 660, eBioscience™ |
| Class | Monoclonal |
| Type | Antibody |
| Clone | B33B4WP |
| Conjugate | eFluor™ 660 |
| Excitation/Emission Max | 651/668 nm |
| Form | Liquid |
| Concentration | 5 µL/Test |
| Purification | Affinity chromatography |
| Storage Buffer | PBS, pH 7.2, with 0.2% BSA |
| Contains | 0.09% sodium azide |
| Storage Conditions | 4°C, store in dark, DO NOT FREEZE |
| Shipping Conditions | Ambient (domestic); Wet ice (international) |
| RRID | AB_2574396 |
Additional Formats:
- eFluor 450
- PE
- PE-Cyanine7
- PerCP-eFluor 710
- Request custom conjugation
Product Specific Information
The B33B4WP monoclonal antibody recognizes human NFκB p65 subunit when serine 529 is phosphorylated.
NFκB (nuclear factor kappa-light-chain-enhancer of activated B cells) is a transcription factor regulating genes involved in proliferation, apoptosis, immunity, and cancer. Functional NFκB is a homo- or heterodimer composed of p65 (RelA), c-Rel, RelB, p50, and p52. Its activity is regulated by IκB inhibitors that sequester NFκB in the cytoplasm.
Activation by TNF-α, IL-1β, LPS, or PMA induces IκB degradation, allowing NFκB nuclear translocation and gene regulation. Phosphorylation of p65 at Ser529 or Ser536 positively regulates NFκB activity.
The B33B4WP antibody detects a 65 kDa band in TNF-α–stimulated HeLa cell lysates.
Applications Reported: Intracellular staining followed by flow cytometric analysis.
Applications Tested: Pre-titrated for intracellular staining and flow cytometry of stimulated human peripheral blood cells. Use 5 µL (0.06 µg) per test (100 µL final volume, 10⁵–10⁸ cells/test).
Recommended Protocols:
- Protocol A: Two-step intracellular (cytoplasmic) proteins
- Protocol C: Two-step fixation/methanol
- Protocol B (nuclear proteins) not recommended
Detailed procedures available in BestProtocols® – Staining Intracellular Antigens for Flow Cytometry.
Fluorochrome Information:
eFluor® 660 is an alternative to Alexa Fluor® 647, excited at 633–647 nm and emitting at 668 nm (Red Laser).
Filtration: 0.2 µm post-manufacturing filtered.
Target Information
NF-κB p65 (RELA gene) is located on human chromosome 11. It forms heterodimers with p50 or p52 and plays a key role in transcriptional regulation of immune and inflammatory responses. Activation involves TNF-α–induced phosphorylation of IKK, leading to IκB degradation and NFκB nuclear translocation. NFκB p65 is essential for immunoglobulin κ gene transcription in mature B cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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