Figure 1. Treatment of differentiated THP-1 cells with LPS stimulates phosphorylation of IKKα at Ser176/180 as detected by the PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit #7073 but does not affect the levels of total IKKα detected by PathScan® Total IKKα Sandwich ELISA Kit #7078. Differentiated THP-1 cells were treated with 1 μg/ml LPS for 10 minutes. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using IKKα Antibody #2682 (left) or Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb #2697 (right) are shown in the bottom figure.
The PathScan® Total IKKα Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IKKα. An IKKα rabbit antibody has been coated onto the microwells. After incubation with cell lysates, IKKα (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, an IKKα mouse detection mAb is added to the captured phospho and nonphospho IKKα protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of total IKKα.
Antibodies in kit are custom formulations specific to kit.
The PathScan® IKKα Sandwich ELISA Kit from Cell Signaling Technology detects endogenous levels of total IKKα protein, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
The NF-κB/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IκB proteins (1-3). Most agents that activate NF-κB do so through a common pathway based on phosphorylation-induced, proteasome-mediated degradation of IκB (3-7). The key regulatory step in this pathway involves activation of a high molecular weight IκB kinase (IKK) complex whose catalysis is generally carried out by three tightly associated IKK subunits. IKKα and IKKβ serve as the catalytic subunits of the kinase and IKKγ serves as the regulatory subunit (8,9). Activation of IKK depends upon phosphorylation at Ser177 and Ser181 in the activation loop of IKKβ (Ser176 and Ser180 in IKKα), which causes conformational changes, resulting in kinase activation (10-13).
Explore pathways + proteins related to this product.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a trademark of Cell Signaling Technology, Inc.