Product Pathways - PathScan ELISA
PathScan® Phospho-IKKβ (Ser177/181) Sandwich ELISA Kit #7080
|7080S||1 Kit (96 assays)||---||In Stock||---|
|7080||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
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|Kit Includes||Volume||Solution Color|
|Phospho-IKKbeta(S177/181) Ab Coated Microwells||96 tests|
|IKKbeta 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|
Note: 12 8-well modules – Each module is designed to break apart for 8 tests.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
The PathScan® Phospho-IKKβ (Ser177/181) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IKKβ when phosphorylated at Ser177/181. A phospho-IKKβ (Ser177/181) rabbit mAb has been coated onto the microwells. After incubation with cell lysates, phospho-IKKβ (Ser177/181) protein is captured by the coated antibody. Following extensive washing, an IKKβ mouse detection mAb is added to detect the captured IKKβ 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 IKKβ phosphorylated at Ser177/181.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
The PathScan® Phospho-IKKβ (Ser177/181) Sandwich ELISA Kit detects endogenous levels of IKKβ protein when phosphorylated at Ser177/181, 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.
ELISA - Western correlation
Figure 1. Treatment of differentiated THP-1 cells with LPS stimulates phosphorylation of IKKβ at Ser177/181 as detected by the PathScan® Phospho-IKKβ (Ser177/181) Sandwich ELISA Kit #7080, but does not affect the level of total IKKβ protein. 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 IKKβ (2C8) Rabbit mAb #2370 (left panel) or Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb #2697 (right panel) 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).
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- 2078 Phospho-IKKα (Ser176)/IKKβ (Ser177) (C84E11) Rabbit mAb
- 2697 Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb
- 2694 Phospho-IKKα/β (Ser176/180) Antibody II
- 2370 IKKβ (2C8) Rabbit mAb
- 8943 IKKβ (D30C6) Rabbit mAb
- 7076 Anti-mouse IgG, HRP-linked 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
For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.