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30 mai 2012
Chargement
Detects and quantifies the level of phosphorylated Hsp27 protein in cell lysates, tissue extracts, and biological fluids. Hsp27 is the small heat shock protein that acts as a molecular chaperone interacting with a large number of proteins. Hsp27 is known for its ability to negatively regulate the mitochondrial pathway to caspase activation. The phosphorylated form of Hsp27 also plays a major role in apoptosis through its ablity to prevent cell death by maintaining redox homeostasis and mitochondrial stability.
| Informations produit | |||
|---|---|---|---|
| Format | 96-well plate | ||
| Form | 96 Tests | ||
| Detection method | Colorimetric | ||
| Assay range | 0.1-10 ng/ml | ||
| Assay time | 4.5 h | ||
| Sample type | Cell lysates | ||
| Positive control | Phosphorylated Hsp27 Standard, UV-treated HeLa or MCF7 cells or sorbitol-treated DU145 cells | ||
| Kit contains | Hsp27-Coated Plate, Rabbit Anti-Hsp27 Detector Antibody, Phosphorylated Hsp27 Standard, HRP-Conjugate, Assay Diluent, ELISA 20X Plate Wash Concentrate, TMB Substrate, ELISA Stop Solution, and a user protocol | ||
| Information relative au stockage et expédition | |||
|---|---|---|---|
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Traitement des matériaux dangereux : En raison de la nature dangereuse des matériaux contenus dans cet envoi, des frais d'expédition supplémentaires pourront être appliqués à votre commande. Certaines tailles peuvent être exonérées des frais d'expédition supplémentaires pour matériaux dangereux. Veuillez contacter la filiale la plus proche pour de plus amples informations sur ces frais. |
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| Catégorie de stockage | -20°C | ||
| Ship |
Blue Ice Only
Regulatory Review |
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| Données | |||
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 A standard curve was generated using phosphorylated (black square) and unphosphorylated (black triangle) Hsp27 as outlined in the Detailed Protocol. Assay Range: 0.1-10 ng/ml; lower limit of detection: ~0.1 ng/ml.  Various tumor cell lines (indicated in the figure) were exposed to 100 J/m2 UV irradiation or treated with 400 mM sorbitol for 30 min. Cell lysates were prepared using CytoBuster™ Protein Extraction Reagent (Cat. No. 71009) according to the recommended protocol. The level of phosphorylated Hsp27 was determined as outlined in the Detailed Protocol and expressed as ng phosphorylated Hsp27/mg total protein.  <b>(A)</b> MCF-7 and HeLa cells were either left untreated (-UV) or exposed to UV-C radiation (+UV). Cell lysates were prepared using CytoBuster™ Protein Extraction Reagent (Cat. No. 71009), subjected to SDS-PAGE, and transferred to nitrocellulose membrane for Western blotting analysis using Anti-Phospho-Hsp27 Detector Antibody Lane 1: MWM; lane 2: MCF-7, -UV; lane 3: MCF-7, +UV; lane 4: HeLa, -UV; lane 5: HeLa, +UV. <b>(B)</b> Unphosphorylated Hsp27 and recombinant Hsp27 that had been phosphorylated <i>in vitro</i> with MAPKAP-2 kinase were subjected to SDS-PAGE and Western blotting analysis using Anti-Phospho-Hsp27 Detector Antibody. Lane 1: MWM; lane 2: in vitro phosphorylated, recombinant Hsp27; lane 3: unphosphorylated, recombinant Hsp27.  MCF7 cells were either left untreated (left bar) or treated with a highly specific, cell-permeable inhibitor of p38 MAP kinase, SB203580 (Cat. No. 559389), (2 µM) followed by UV treatment (right bar). Cell lysates were prepared using CytoBuster™ Protein Extraction Reagent (Cat. No. 71009) and the level of phosphorylated Hsp27 was determined as outlined in the Detailed Protocol. 
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