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Merck KGaA
Frankfurter Str. 250
64293 Darmstadt
Germany
Telefon: +49 6151 72-0
Fax: +49 6151 72 2000
15 Februar 2012
wird geladen
Detects and quantifies the level of IκBα protein phosphorylated at Ser32 in human cells. IκBα belongs to IκB protein family and is known to modulate NF-kB (nuclear factor-κB) nuclear translocation. It plays an important role in immune and inflammatory responses, cell division and apoptosis.
| Produktinformationen | |||
|---|---|---|---|
| Format | 96-well plate | ||
| Form | 96 Tests | ||
| Detection method | Colorimetric | ||
| Species reactivity | human, not mouse, not rat | ||
| Unit definition | One unit is defined as the amount of IκBα pSer32 derived from 40 pg IκBα phosphorylated by IKKα. | ||
| Sensitivity | < 1.0 Units/ml | ||
| Assay range | 1.6-100 Units/ml | ||
| Assay time | 4 h | ||
| Kit contains | IκBα (pSer32) Standard, Standard Diluent Buffer, IκBα Antibody-Coated 96-Well Plate, Rabbit Anti-IκBα (pSer32) Detector Antibody, Anti-Rabbit IgG-HRP Concentrate, HRP Diluent, Wash Buffer Concentrate, TMB, Stop Solution, Plate Covers and a user protocol. | ||
| Lager- und Versandinformationen | |||
|---|---|---|---|
| Lagerkategorie | +2°C to +8°C | ||
| Ship |
Blue Ice Only
Multiple Toxicity Values, refer to MSDS |
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| Daten | |||
|---|---|---|---|
 Known quantities of IκBα (pSer<sup>32</sup>) protein were measured as outlined in the Detailed Protocol and by immunoblotting using an anti-IκBα (pSer<sup>32</sup>) primary antibody and chemiluminescent detection. The ELISA is approximately 2x more sensitive than immunoblotting.  The specificity of this assay for IκBα phosphorylated at Ser<sup>32</sup> was confirmed by peptide competition. The data show that only the phosphopeptide containing the phosphorylated Ser<sup>32</sup> could block the ELISA signal in Jurkat cells treated with TNF-α. The same sequence containing non-phosphorylated serine at position 32 did not block the signal. A peptide conatining a phosphoserine at position 36 also did not block the signal.  Cell lysates made from Jurkat cells were harvested at various times following treatment with TNF-α and compared to non-treated controls, as determined using the Detailed Protocol provided. The data presented here also demonstrate that there is an initial increase in phosphorylation followed by a reduction in the level of Total IκBα which is attributable to the TNF-α-stimulated degradation of IκBα. The results correlate well with immunoblot analysis.   Samples of known IκBα (pSer<sup>32</sup>) concentration were assayed in replicates of 16 to determine precision within an assay.  Samples were assayed 48 times in multiple assays to determine precision between assays. |
