Product Pathways - PathScan ELISA
PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit #7073
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
|Kit Includes||Volume||Solution Color|
|Phospho-IKK(S176/180) Ab Coated Microwells||96 tests|
|IKKalpha Detection Ab||11 ml||Green|
|Anti-mouse IgG, HRP-linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|PathScan® Sandwich ELISA Lysis Buffer (1X) #7018||30 ml||Colorless|
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
The PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IKKα when phosphorylated at Ser176/180. A phospho-IKKα (Ser176/180) rabbit mAb has been coated onto the microwells. After incubation with cell lysates, phospho-IKKα (Ser176/180) protein is captured by the coated antibody. Following extensive washing, an IKKα mouse detection mAb is added to detect the captured 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 the developed color is proportional to the quantity of IKKα phosphorylated at Ser176/180.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
The PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit from Cell Signaling Technology detects endogenous levels of IKKα protein when phosphorylated at Ser176/180, 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.
Figure 2. The relationship between the protein concentration of lysates from untreated and LPS-treated THP-1 cells and the absorbance at 450 nm using the PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit is shown.
ELISA - Western correlation
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 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).
- Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-6.
- Beg, A.A. and Baldwin, A.S. (1993) Genes Dev 7, 2064-70.
- Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8.
- Brown, K. et al. (1995) Science 267, 1485-8.
- Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18.
- Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
- Chen, Z.J. et al. (1996) Cell 84, 853-62.
- Zandi, E. et al. (1997) Cell 91, 243-52.
- Karin, M. (1999) Oncogene 18, 6867-74.
- DiDonato, J.A. et al. (1997) Nature 388, 548-54.
- Mercurio, F. et al. (1997) Science 278, 860-6.
- Johnson, L.N. et al. (1996) Cell 85, 149-58.
- Delhase, M. et al. (1999) Science 284, 309-13.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
- 7078 PathScan® Total IKKα Sandwich ELISA Kit
- 2078 Phospho-IKKα (Ser176)/IKKβ (Ser177) (C84E11) Rabbit mAb
- 2694 Phospho-IKKα/β (Ser176/180) Antibody II
- 2697 Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb
- 2682 IKKα Antibody
- 7018 PathScan® Sandwich ELISA Lysis Buffer (1X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 9998 BSA
- 7004 TMB Substrate
- 7002 STOP Solution
Rabbit monoclonal antibody is produced under license (granting certain rights including those under U. S. Patents No. 5,675,063 and 7,429,487) from Epitomics, Inc.
For Research Use Only. Not For Use In Diagnostic Procedures.