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Citations Search

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J. Biol. Chem. April 28, ePub ahead of print. Mitogenic responses of vascular smooth muscle cells to lipid peroxidation-derived aldehyde 4-hydroxy-trans-2-nonenal (HNE): Role of aldose reductase-catalyzed reduction of the HNE-glutathione conjugates in regulating cell growth. 2006

Ramana, K.V., Bhatnagar, A., Srivastava, S., Umesh C.S. Yadav, Awasthi, S., Awasthi, Y.C. and Srivastava, S.K.

Notes: Protein Kinase C activity in rat vascular smooth muscle cells in response aldose reductase was measured using the SignaTECT® Protein Kinase C (PKC) Assay System. Expression of NFκB and AP-1 was investigated by electrophoretic mobility gel shift assays using the AP-1 and NFκB Consensus Oligonucleotides. (3406)

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Nucl. Acids Res. 29, 21. Development of a sensitive multi-well colorimetric assay for active NF-κB. 2001

Renard, P., Ernest, I., Houbion, A., Art, M., Le Calvez, H., Raes, M. and Remacle, J.

Notes: A double-stranded NF-κB consensus oligonucleotide sequence containing 122bp oligonucleotide was created by PCR with a biotinylated primer at the 5´ end and the NF-κB Consensus Oligonucleotide sequence at the 3´ end. The resultant probe was bound to streptavidin-coated plates and used to screen for activated NF-κB in IL-1β-stimulated WI-38 VA13 human fibroblast cell lysates. Actively bound NF-κB was detected colorimetrically with either a rabbit anti NF-kB p50 or p65 antibody, followed by a goat anti-rabbit HRP conjugated secondary antibody and a TMB solution. Recombinant hNF-κB (p50) was used as a standard in the assay. The authors provide details for each step in the process including the specific buffers, lysate volumes, and incubation times and temperatures. The NF-κB Consensus Oligonucleotide was also used in EMSAs to confirm the results observed in the screening assay. For this assay the authors used 25μg of total protein from each lysate preparation. (2690)

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J. Biol. Chem. 273, 6607-6610. IκBα degradation and nuclear factor-κB DNA binding are insufficient for interleukin-1β and tumor necrosis factor-α-induced κB-dependent transcription. Requirement for an additional activation pathway. 1998

Bergmann, M., Hart, L., Lindsay, M., Barnes, P.J. and Newton, R.

Notes: IL-1β and TNFα  have been shown to induce two p50/p65 NF-κB DNA binding complexes in A549 cells. Tfx™-50 Reagent was used to stably transfect A549 cells in order to study the effects on NF-κB-dependent transcription. Promega's Luciferase Assay System was used to measure the effect of kinase inhibitors on IL-1β and TNFα stimulated NF-κB-dependent transcription. In addition, Promega's NF-κB and OCT1 Oligonucleotides were used in gel shift assays (1438)

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J. Biol. Chem. 273, 32787-32792. Protein Kinase C-alpha modulates lipopolysaccharide-induced functions in a murine macrophage cell line. 1998

St Denis, A., Chano, F., Tremblay, P., St Pierre, Y., and Descoteaux, A.

Notes: The authors examined the effect of overexpressing a dominant negative mutant of protein kinase C alpha in RAW 264.7 cells on LPS induced signal transduction. They used the Anti-ACTIVE® JNK pAb for western analysis and the NF-kappaB oligonucleotide for electrophoretic mobility shift assay (EMSA) analysis of NF-kappaB activation. (0326)

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