Product Pathways - PathScan ELISA
PathScan® Total IKKα Sandwich ELISA Kit #7078
|7078S||1 Kit (96 assays)||---||In Stock||---|
|7078||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at email@example.com.
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|Kit Includes||Volume||Solution Color|
|IKKalpha Ab Coated Microwells||96 assays|
|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.
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® 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
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.
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 PathScan® Total IKKα 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).
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- 7073 PathScan® Phospho-IKKα (Ser176/180) Sandwich ELISA Kit
- 11930 IKKα (3G12) Mouse mAb
- 2682 IKKα Antibody
- 2697 Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb
- 2694 Phospho-IKKα/β (Ser176/180) Antibody II
- 2078 Phospho-IKKα (Ser176)/IKKβ (Ser177) (C84E11) Rabbit mAb
- 9803 Cell Lysis Buffer (10X)
- 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.