WB result of Phospho-NAK/TBK1 (Ser172) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho-NAK/TBK1 (Ser172) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa starve overnight, then treated with 100 nM Calyculin A for 30 minutes whole cell lysate 20 µg
Lane 3: untreated Jurkat whole cell lysate 20 µg
Lane 4: Jurkat treated with 100 nM Calyculin A for 30 minutes whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 84 kDa
Observed MW: 84 kDa
Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Phospho-NAK/TBK1 (Ser172) |
| Synonyms | Serine/threonine-protein kinase TBK1; NF-kappa-B-activating kinase; T2K; TANK-binding kinase 1; NAK; TBK1 |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm |
| Accession | Q9UHD2 |
| Clone Number | S-2481-14 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC |
| Reactivity | Hu, Ms, Rt |
| Predicted Reactivity | Xe |
| 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 |
| Dot Blot | 1:1000 | |
| WB | 1:1000-1:10000 | Hu, Ms, Rt |
| ICC | 1:100 | Hu |
Background
Phospho-NAK/TBK1 (Ser172) protein is a crucial component in cellular signaling pathways. NAK, also known as TBK1, is a serine/threonine kinase that plays a significant role in innate immune responses and NF-κB activation. When phosphorylated at serine 172, it becomes activated and can phosphorylate downstream substrates such as IRF3, leading to the production of type I interferons. This phosphorylation event at Ser172 is essential for its kinase activity and its involvement in antiviral signaling and inflammatory responses. Additionally, TBK1 has been implicated in autophagy regulation through its interaction with other proteins, further highlighting its multifaceted functions in maintaining cellular homeostasis and responding to various stressors.
Picture
Picture
Western Blot
WB result of Phospho-NAK/TBK1 (Ser172) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho-NAK/TBK1 (Ser172) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated RAW264.7 whole cell lysate 20 µg
Lane 2: RAW264.7 treated with 100 nM Calyculin A for 30 minutes whole cell lysate 20 µg
Lane 3: untreated NIH/3T3 whole cell lysate 20 µg
Lane 4: NIH/3T3 treated with 100 nM Calyculin A for 30 minutes whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 84 kDa
Observed MW: 84 kDa
WB result of Phospho-NAK/TBK1 (Ser172) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho-NAK/TBK1 (Ser172) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated C6 whole cell lysate 20 µg
Lane 2: C6 starve overnight, then treated with 100 nM Calyculin A for 30 minutes whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 84 kDa
Observed MW: 84 kDa
Dot Blot
Dot blot result of Phospho-NAK/TBK1 (Ser172) Recombinant Rabbit mAb
Lane 1: NAK/TBK1 (Ser172) phospho peptide
Lane 2: NAK/TBK1 (Ser172) unmodified peptide
Primary antibody: Phospho-NAK/TBK1 (Ser172) Recombinant Rabbit mAb at 1/1000 dilution
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Immunocytochemistry
ICC analysis of HeLa cells serum starvation (overnight), then treated with Calyculin A (100nM, 30 min) (top panel) and untreated HeLa cells (below panel). Anti- Phospho-NAK/TBK1 (Ser172) antibody was used at 1/100 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
