Cellomics HCS Reagent Kit for detection of active heme oxygenase 1. | | | | | normal heme
oxygenase 1
expression | treat cells | induced heme
oxygenase 1 | analyze | conclude |
DescriptionThe Cellomics Heme Oxygenase 1 Activation Kit measures activation of heme oxygenase 1 in cells. The kit contains a mouse monoclonal heme oxygenase 1 primary antibody, a DyLight 549-Conjugated Secondary Antibody, and various other reagents and buffers required for immunofluorescent detection for high-content screening (HCS) assays. Heme oxygenase 1 is a microsomal enzyme that catalyses the oxidation of heme to antioxidants, biliveridin and carbon monoxide and protects cells from wide variety of stress conditions through activation of p38 MAPK. Carbon monoxide produced during oxidation of heme by heme oxygenase 1 activates p38 MAPK, which confers tissue protection through inhibition of cytokine production. Heme oxygenase 1 can be induced by oxidative stress, hypoxia, heat shock, heavy metals and cytokines. To measure heme oxygenase 1 levels, A549 cells were treated with copper sulfate. In normal cells, heme oxygenase 1 protein levels are low. Upon activation, heme oxygenase 1 increases in the cytoplasm and nucleus, as evidenced by increased staining (Figure 1). The assay was developed in A549 cells and optimized using the Thermo Scientific ArrayScan HCS Reader and Compartmental Analysis Bioapplication Software Module. Stained cells also can be imaged using fluorescence or confocal microscopy. | Figure 1. Staining of Heme oxygenase 1 in A549 cells treated with or without 0.5 mM copper sulfate for 24 hours. Cells were stained according to the kit protocol and imaged using the ArrayScan HCS Reader. |
| Figure 2. Dose response curve of heme oxygenase 1 in A549 cells treated with copper sulfate for 24 hours. Data represents mean ±SD from three plates (eight wells per 96-well plate per dose of copper sulfate). |
Related Poster Presentations | Kay Opperman, Bhaskar S. Mandavilli, and Richik N. Ghosh (2008) Assessment of Oxidative Stress and DNA Damage in Individual Cells Using Automated, Quantitative Cell-Based Imaging. |
Kit Contents| Component | 8405801 | 8405802 | | Heme Oxygenase 1 Primary Antibody | 15 µl | 75 µl | | DyLight 549 Conjugated Goat Anti-Mouse IgG | 30 µl | 72 µl | | Hoechst Dye | 30 µl | 30 µl | | Wash Buffer (10X Dulbecco's PBS) | 100 ml | 100 ml | | Wash Buffer (10X Dulbecco's PBS with Tween®-20) | 100 ml | 100 ml | Permeabilization Buffer (10X Dulbecco's PBS
with 1% Triton® X-100) | 100 ml | 100 ml | | Blocking Buffer (10X) | 85 ml | 85 ml | | Thin Plate Seal Assembly | 7/pack | 7/pack |
Ordering InformationScreening size kits and components (bulk and custom) available with special pricing. Please call. To order Cellomics HCS Reagent Kits, please call 800-874-3723 in the U.S.
or contact your local distributor of Thermo Scientific Products. Choose "Order Online" in table to see pricing and order online (U.S. only). | Buy | Product # | Description | Pkg. Size | | Order Online | 8405801 | Heme Oxygenase 1 Activation Kit | 1 x 96 | | Order Online | 8405802 | Heme Oxygenase 1 Activation Kit | 5 x 96 |
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