Nuclear NF-κB expression was elevated 30-min following LPS treatment, maximal by 45-min, and returned to basal levels by 24-h. Binding of nuclear NF-κB to consensus oligonucleotide sequences followed a similar pattern to that observed for p-NF-κB, but lasted slightly longer.

NF-κB p65 是NF-κB家族成员之一，NF-κB是一种多效性转录因子，几乎存在于所有细胞类型中，并参与许多生物学过程，例如炎症、免疫、分化、细胞生长、肿瘤发生和凋亡。 NF-κB受翻译后修饰和亚细胞区室化的各种机制以及与其他辅因子或共抑制因子的相互作用的控制。 p65-RelA是哺乳动物NF-κB家族的成员，该家族由5个亚基组成：p50-NF-κB1，p52-NF-κB2，c-Rel，p65-RelA和RelB。

2021年3月1日 · SNX25 knockdown decreased cytoplasmic p65 after 30 min LPS stimulation (SNX25 siRNA, 0.74 ± 0.19-fold control P = 0.012). In addition, SNX25 knockdown increased nuclear p65 though the difference was not statistically significant (SNX25 siRNA, 1.50 ± 0.84-fold control P = 0.15) ( Fig 4C ).

Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Immunohistochemical analysis of paraffin-embedded Human-uterus tissue. 1,NFκB-p65 (phospho Ser536) Polyclonal Antibody was diluted at 1:200(4°C,overnight). 2, Sodium citrate pH 6.0 was used for antibody retrieval(>98°C,20min). 3,Secondary antibody was diluted at 1:200(room tempeRature, 30min). Negative control was used by secondary antibody ...

LPS作用30min后，Western blotting 检测不同实验组细胞核、细胞质内NF-κB p65蛋白水平，LPS作用9h后，实时荧光定量PCR法检测NF-κB p65、COX-2 mRNA水平。 结果 LPS作用30min后，各实验组细胞NF-κB p65蛋白总量差异不显著，但LA和PDTC预处理组细胞核内NF-κB...

2025年3月4日 · A previous study showed that p65 phosphorylation at Ser 276 facilitated its interaction with p300/CREB (adenosine 3′,5′-monophosphate response ... for 30 min, and the extracted cytosolic proteins were followed by immunoprecipitation with anti-GCN5 or anti-p65 antibodies. The interaction between GCN5 and NF-κB p65 was analyzed by WB assay ...

1 天前 · NF-κB p65 remains inactive in the cytoplasm until activated, upon which it is rapidly imported into the nucleus by importin family proteins. ... The mixture was vortexed and incubated for 30 min at 30 °C, followed by centrifugation (12 000 rcf, 5 min). The supernatant was removed, and the pellet was resuspended in 0.01× PBS with 0.1% Tween ...

之前做0，30，60min也没跑出来，可能时间长了磷酸化已经结束了，后来一直跑的0，15，30min，多试几个时间点

2024年10月30日 · The nuclear translocation of p65 was detected using immunofluorescence staining. After washing with PBS, Raw264.7 cells in each group was successively fixed with 4% paraformaldehyde at room temperature for 30 min, permeabilized with Triton X-100 for 15 min, blocked with 5% bovine serum albumin for 2 h.