PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300
12 months from date of receipt / reconstitution, -20 °C as supplied
| 应用 | 稀释度 | 推荐种属 |
|---|---|---|
| Dot Blot | 1:1000 | |
| WB | 1:1000 | Hu,Ms,Rt |
| IP | 1:50 | Hu |
Phospho-NF-κB p65 (Ser468) refers to the phosphorylation of the p65 subunit of NF-κB at serine 468. This specific phosphorylation event plays a distinct role in the regulation of NF-κB signaling. Unlike many other phosphorylation sites on p65 that generally enhance its transcriptional activity, phosphorylation at Ser468 has been shown to inhibit p65's transactivation potential and can be involved in p65 ubiquitination and degradation, thereby reducing its transcriptional activity. This modification is part of the complex post-translational regulation of NF-κB, which is a critical mediator of cellular responses to various stimuli, including inflammation, immune responses, and cellular stress. The phosphorylation status of p65 at Ser468 can impact the cell's inflammatory response and may have implications for diseases where NF-κB signaling is dysregulated.
WB result of Phospho-NF-κB p65 (Ser468) Recombinant Rabbit mAb
Primary antibody: Phospho-NF-κB p65 (Ser468) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa treated with 20 ng/ml TNF-alpha and 50 nM Calyculin A for 5 minutes whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 60 kDa
Observed MW: 70 kDa
WB result of Phospho-NF-κB p65 (Ser468) Recombinant Rabbit mAb
Primary antibody: Phospho-NF-κB p65 (Ser468) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: Untreated NIH/3T3 whole cell lysate 20 µg
Lane 2: NIH/3T3 starve for 3hr then treated with Calyculin A (100nM/ml, 30min) whole cell lysate 20 µg
Negative control: Untreated NIH/3T3 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 60 kDa
Observed MW: 70kDa
WB result of Phospho-NF-κB p65 (Ser468) Recombinant Rabbit mAb
Primary antibody: Phospho-NF-κB p65 (Ser468) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: Untreated C6 whole cell lysate 20 µg
Lane 2: C6 starve overnight then treated with TNF-α (20ng/ml, 30min) and Calyculin A (100nM/ml, 30min) whole cell lysate 20 µg
Negative control: Untreated C6 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 60 kDa
Observed MW: 70kDa
Phospho-NF-κB p65 (Ser468) Recombinant Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating phospho-NF-κB p65 (Ser468) in 0.4 mg HeLa + TNF-alpha (20 ng/ml) and Calyculin A (50 nM) for 5 mins whole cell lysate.
Western blot was performed on the immunoprecipitate using Phospho-NF-κB p65 (Ser468) Recombinant Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: HeLa + TNF-alpha (20 ng/ml) and Calyculin A (50 nM) for 5 mins whole cell lysate 20 µg (Input)
Lane 2: Phospho-NF-κB p65 (Ser468) Recombinant Rabbit mAb IP in HeLa + TNF-alpha (20 ng/ml) and Calyculin A (50 nM) for 5 mins whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in HeLa + TNF-alpha (20 ng/ml) and Calyculin A (50 nM) for 5 mins whole cell lysate
Predicted MW: 60 kDa
Observed MW: 70 kDa
Exposure time: 超敏90 s
Dot blot result of Phospho-NF-κB p65 (Ser468) Recombinant Rabbit mAb
Lane 1: NF-κB p65 (Ser468) phospho peptide
Lane 2: NF-κB p65 unmodified peptide
Primary antibody: Phospho-NF-κB p65 (Ser468) Recombinant Rabbit mAb at 1/1000 dilution
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
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