WB result of Phospho-NF-κB p65 (Ser536) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody incubation conditions: overnight at 4°C
Primary antibody: Phospho-NF-κB p65 (Ser536) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa treated with 100 ng/ml Calyculin A for 30 minutes and 20 ng/ml TNF-α 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
Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Phospho-NF-κB p65 (Ser536) |
| Synonyms | Transcription factor p65; Nuclear factor NF-kappa-B p65 subunit; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3; NFKB3 |
| Location | Cytoplasm, Nucleus |
| Accession | Q04206 |
| Clone Number | S-3597 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB |
| Reactivity | Hu, Ms, Rt, Mk, Hm |
| Positive Sample | HeLa (treated with 100 ng/ml Calyculin A for 30 minutes and 20 ng/ml TNF-α for 5 minutes) |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000-1:2000 | Hu, Ms, Rt, Hm, Mk, Pg |
Background
Phospho-NF-κB p65 (Ser536) refers to the phosphorylation of the p65 subunit (RelA) of the NF-κB transcription factor complex at serine residue 536, a key post-translational modification that significantly enhances its transcriptional activity. This phosphorylation is primarily mediated by IκB kinases (IKKα and IKKβ) as well as other kinases such as TBK1 and RSK in response to pro-inflammatory stimuli like TNF-α, IL-1, or lipopolysaccharide (LPS). Phosphorylation at Ser536 promotes the nuclear translocation of p65, facilitates its interaction with co-activators such as CBP/p300, and increases its binding affinity to specific DNA κB sites, thereby amplifying the expression of target genes involved in inflammation, immune responses, cell survival, and proliferation. Due to its central role in activating the canonical NF-κB pathway, Phospho-p65 (Ser536) is widely utilized as a critical biomarker for monitoring inflammatory signaling activation in various physiological and pathological contexts, including cancer, autoimmune diseases, and chronic inflammation.
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Western Blot
