Product Includes | Product # | Quantity | Color | Storage Temp |
---|---|---|---|---|
IKKα Mouse Detection mAb | 13712 | 1 ea |
|
+4C |
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) | 13304 | 1 ea |
|
+4C |
Detection Antibody Diluent | 13339 | 11 ml |
|
+4C |
HRP Diluent | 13515 | 11 ml |
|
+4C |
TMB Substrate | 7004 | 11 ml |
|
+4C |
STOP Solution | 7002 | 11 ml |
|
+4C |
ELISA Wash Buffer (20X) | 9801 | 25 ml |
|
+4C |
ELISA Sample Diluent | 11083 | 25 ml |
|
+4C |
Sealing Tape | 54503 | 2 ea |
|
+4C |
PathScan® Sandwich ELISA Lysis Buffer (1X) | 7018 | 30 ml |
|
-20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
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.
Specificity/Sensitivity
Background
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.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
Limited Uses
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