Phospho-p65 Antibody (pS529)

Overview

Phospho-p65 Antibody (pS529) is a research-use antibody directed against PHOSPHO-P65. It is supplied for use in common immunoassay contexts such as WB (RUO).

Key elements and design rationale

• Target: PHOSPHO-P65.
• Description (provided): types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis.
• Antibody type: Rabbit, clone AbN84, Rabbit IgG.
• Format: Purified; Protein A affinity.
• Reported/predicted localization: Nucleus, Cytoplasm.
• Species reactivity: tested: Human.
• Immunogen (if provided): The amino acids surrounding a phosphorylated Serine at position 529 were used as the immunogen for the phospho-p65 antibody..

The information above helps you match the antibody format to your assay context, interpret species-dependent differences, and anticipate how epitope context (isoforms, PTMs, or conformational state) may influence signal.

Biological background

NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells. [UniProt]

For curated annotations (gene/protein naming, domains, isoforms, and pathway links) for PHOSPHO-P65, consult primary databases such as UniProt, NCBI Gene, and Ensembl.

Research relevance and current trends

• Context-dependent expression studies: researchers often examine PHOSPHO-P65 abundance and localization across perturbations (genetic, pharmacologic, or environmental) to connect phenotype to molecular changes.
• Reagent reproducibility: there is growing emphasis on antibody specificity checks using orthogonal approaches (e.g., genetic perturbation or independent antibodies) and transparent reporting of clone/lot information.
• Multi-modal datasets: antibody-based readouts are increasingly combined with transcriptomics and imaging to relate protein-level measurements to cell-state transitions.

Common research applications

• Western blotting (immunoblot) for relative detection of target protein abundance and apparent molecular weight.

When comparing conditions, interpret changes in signal in the context of sample composition, expected localization, and any known isoform complexity for the target.

Notes for experimental interpretation

• Isoforms and PTMs: alternative splicing or post-translational modifications can change epitope accessibility and apparent molecular weight; interpret bands/signals accordingly.
• Cross-reactivity and matrix effects: background binding can vary by sample type, species, and blocking/detection chemistries; include appropriate negative controls.
• Control concepts: where feasible, use genetic perturbation (KO/KD/overexpression), orthogonal assays, or independent antibodies to support specificity claims.

Antibody considerations: Polyclonal reagents may recognize multiple epitopes and can increase sensitivity but may show broader binding profiles, while monoclonal clones provide a single-epitope readout that can improve consistency across experiments. If a conjugate is listed, the antibody supports more direct detection workflows; otherwise, it is typically used with a compatible secondary antibody.

Customization & Add-ons: Can’t find the antibody you need—or require a custom format for your assay? We can help you source the best match or support custom antibody solutions for diverse research needs, including species and isotype selection, conjugations and labeling (e.g., HRP/AP, biotin, fluorophores), purification grade options (Protein A/G, affinity purified), formulation preferences (buffer selection, carrier-free, glycerol-free), custom concentrations and aliquoting, low-endotoxin options for cell-based work, and application-focused QC/validation support (project dependent). Click Talk to a Scientist to submit a request, email us at support@biohippo.com, or explore our Research Services for additional support—our team will follow up with feasibility details and next steps.