Phospho-NF-κB p65 (Ser529) Antibody [F5A1]

Catalog No.: F4238

Application: Reactivity: Human

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)

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代表番号: 045-509-1970|電子メール：sales@selleck.co.jp

使用情報

Dilution
WB1:1000 IP1:50 IF1:3200 FCM1:100-1:400

Application
WB, IP, IF, FCM

Source
Rabbit Monoclonal Antibody

Reactivity
Human

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

Predicted MW
65 kDa

Datasheet & SDS

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.

Clone
F5A1

Synonym(s)
Transcription factor p65; Nuclear factor NF-kappa-B p65 subunit; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3; RELA; NFKB3

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.

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.

References
[1] Maguire O, et al. J Immunol Methods. 2015 Aug;423:3-11. [2] Giridharan S, et al. J Inflamm Res. 2018 Oct 30;11:407-419.

PVDFメンブレン孔径参考表

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%