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PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit #7355

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ELISA Image 1: PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit
Figure 1: Treatment of HeLa cells with TNF-α stimulates phosphorylation of IκBα at Ser32, detected by PathScan® Phospho-IκBα (Ser32) Sandwich ELISA kit, #7355, but does not affect the level of total IκBα protein detected by PathScan® Total IκBα Sandwich ELISA kit, #7360. Treatment with MG132, a proteasome inhibitor (37ºC for 180 min before TNF-α induction), causes accumulation of phospho-IκBα in control and TNF-α-treated cells, shown in both Sandwich ELISA and Western blot analysis. OD 450 readings are shown in the top figure, while the corresponding Western blot using Phospho-IκBα (Ser32) Ab #9241 (right panel) or IκBα (L27H11) Mouse mAb #7361 (left panel), is shown in the bottom figure.
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Custom Ordering Details:

If kit quantities from the same lot are needed in unlisted sizes, contact us for processing time and pricing.

Looking for this ELISA kit in a 384-well format? Inquire for availability, processing time, and pricing.

Supporting Data

REACTIVITY H M
Application Key:
  • ELISA-ELISA 
Species Cross-Reactivity Key:
  • H-Human 
  • M-Mouse 
  • Product Includes
  • Related Products
Product IncludesVolumeSolution Color
IkappaBα Mouse mAb Coated Microwells #2421896 tests
Phospho-IkBa (Ser32) Rabbit Detection mAb #141221 eaGreen (Lyophilized)
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) #132721 eaRed (Lyophilized)
Detection Antibody Diluent #1333911 mlGreen
HRP Diluent #1351511 mlRed
TMB Substrate #700411 ml
STOP Solution #700211 ml
Sealing Tape #545032 ea
ELISA Wash Buffer (20X) #980125 ml
ELISA Sample Diluent #1108325 mlBlue
Cell Lysis Buffer (10X) #980315 ml

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Product Information

Product Description

CST's PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Phospho-IκBα (Ser32) protein. An IκBα Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both nonphospho- and phospho-IκBα proteins are captured by the coated antibody. Following extensive washing, Phospho-IκBα (Ser32) Antibody is added to detect the captured phospho-IκBα (Ser32) protein. Anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of phospho-IκBα (Ser32) protein.

*Antibodies in kit are custom formulations specific to kit.

Protocol

Specificity / Sensitivity

CST's PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit detects endogenous levels of Phospho-IκBα (Ser32) protein. As shown in Figure 1, using this Sandwich ELISA Kit #7355, a significant induction of Phospho-IκBα (Ser32) in HeLa cells treated with TNF-α can be detected. However, the level of total IκBα (phospho- and nonphospho-), detected by the Total IκBα Sandwich ELISA Kit #7360, remains unchanged. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Species Reactivity:

Human, Mouse

Background

The NF-κB/Rel transcription factors are present in the cytosol in an inactive state complexed with the inhibitory IκB proteins (1-3). Activation occurs via phosphorylation of IκBα at Ser32 and Ser36 followed by proteasome-mediated degradation that results in the release and nuclear translocation of active NF-κB (3-7). IκBα phosphorylation and resulting Rel-dependent transcription are activated by a highly diverse group of extracellular signals including inflammatory cytokines, growth factors, and chemokines. Kinases that phosphorylate IκB at these activating sites have been identified (8).

Pathways

Explore pathways related to this product.


Database Links

UniProt ID: P25963


Entrez-Gene Id: 4792


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