Datasheet

生物学的記述

Specificity	Phospho-NF-κB p65 (Ser529) Antibody [F5A1] detects endogenous levels of total NF-κB p65 protein only when it is phosphorylated at Ser529.
Background	Phospho-NF-κB p65 at Ser529 (p65-S529ph) is a post-translational modification of the transcriptionally active p65 subunit within the canonical NF-κB pathway, modulating DNA-binding affinity, promoter engagement, and gene transcription. In resting cells, p65 is sequestered in the cytoplasm by IκB proteins. Upon cytokine stimulation (e.g., TNF-α), the IKK complex phosphorylates and degrades IκB, freeing p65-containing dimers for nuclear translocation and binding to κB-motif promoters of genes controlling inflammation, cell survival, and immunity. Ser529 phosphorylation, mediated by casein kinase II and related kinases after IκB dissociation, occurs within the p65 transactivation domain. While this modification does not impede nuclear entry, it enhances p65’s ability to recruit coactivators and chromatin-modifying complexes, thereby amplifying transcriptional activation of NF-κB-dependent pro-inflammatory and anti-apoptotic genes. P65-S529ph signaling contributes to cytokine-driven endothelial activation, macrophage polarization, and T- and B-cell immune responses, making this modification a valuable readout for NF-κB pathway engagement in both innate and adaptive immunity. Dysregulation of Ser529 phosphorylation in chronic inflammation and cancer leads to persistent NF-κB activity, resistance to apoptosis, and pro-tumorigenic signaling, implicating aberrant p65-S529ph in various inflammatory and oncogenic diseases.

Specificity

Phospho-NF-κB p65 (Ser529) Antibody [F5A1] detects endogenous levels of total NF-κB p65 protein only when it is phosphorylated at Ser529.

Background

Phospho-NF-κB p65 at Ser529 (p65-S529ph) is a post-translational modification of the transcriptionally active p65 subunit within the canonical NF-κB pathway, modulating DNA-binding affinity, promoter engagement, and gene transcription. In resting cells, p65 is sequestered in the cytoplasm by IκB proteins. Upon cytokine stimulation (e.g., TNF-α), the IKK complex phosphorylates and degrades IκB, freeing p65-containing dimers for nuclear translocation and binding to κB-motif promoters of genes controlling inflammation, cell survival, and immunity. Ser529 phosphorylation, mediated by casein kinase II and related kinases after IκB dissociation, occurs within the p65 transactivation domain. While this modification does not impede nuclear entry, it enhances p65’s ability to recruit coactivators and chromatin-modifying complexes, thereby amplifying transcriptional activation of NF-κB-dependent pro-inflammatory and anti-apoptotic genes. P65-S529ph signaling contributes to cytokine-driven endothelial activation, macrophage polarization, and T- and B-cell immune responses, making this modification a valuable readout for NF-κB pathway engagement in both innate and adaptive immunity. Dysregulation of Ser529 phosphorylation in chronic inflammation and cancer leads to persistent NF-κB activity, resistance to apoptosis, and pro-tumorigenic signaling, implicating aberrant p65-S529ph in various inflammatory and oncogenic diseases.

使用情報

Application

WB, IP, IF, FCM

Dilution

WB	IP	IF	FCM
1:1000	1:50	1:3200	1:100-1:400

Reactivity

Human

Source

Rabbit Monoclonal Antibody

MW

65 kDa

Storage Buffer

PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage
(from the date of receipt)

-20°C (avoid freeze-thaw cycles), 2 years

プロトコール

References

https://pubmed.ncbi.nlm.nih.gov/25862606/
https://pubmed.ncbi.nlm.nih.gov/30464573/

Application Data

WB

Validated by Selleck

Lane 1: THP-1 (TPA, 50 ng/mL，18 h; rested 48 h), Lane 2: THP-1 (TPA, 50 ng/mL，18 h; rested 48 h; LPS, 5 μg/mL，15 min)