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Cytoskeleton/MICAL-1 Protein 6xHis/2 x 50 ug/MIC01
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DetailsMICAL (MIC01) is an intracellular flavoprotein monoxygenase, conserved from insects to mammals, that functions as a catalyst for oxidation-reduction (redox) reactions. Terman’s group showed that MICAL interacts with F-actin and uses NADPH as a cofactor to oxidize actin at Met44 and Met47. As part of the MOXtrue™ product line, Human MICAL1 protein (MIC01) [accession # Q8TDZ2 (MICA1_HUMAN)] has been produced and purified from a bacterial expression system. The recombinant protein is N-terminally 6x Histidine tagged and is comprised of the Redox and CH domains (aa: 1-612) of the full length protein. Validation studies have shown that MIC01 can oxidize actin which will reduce subtilisin A cleavage by >90% in limited proteolysis assays. MIC01 has also been validated for the production of oxidized actin which can be used in downstream applications such as sedimentation assays and actin polymerization assays. To learn more about using MICAL as a research tool see our Datasheet To learn more about the MICAL/MsrB/Actin physiological redox system see our Newsletter Each lot of purified protein is quality controlled to provide high batch to batch consistency, see COA documents. Validated Applications MICAL-1 (MIC01) Protein Purity DeterminationA 10 μg sample of MICAL-1 protein was sepa-rated by electrophoresis in a 4- 20% Tris-Glycine gel and stained with Coomassie Blue. Protein quantitation was performed using the Precision RedTM Protein Assay Reagent (Cat.# ADV02). Mark12 standard molecular weight markers are from Invitrogen.Subtilisin Assay on MsrB2 Treated Oxidized ActinMICAL-oxidized actin (MetO Actin) (Cat# MXA95) was diluted to 0.1 mg/ml (2.3 μM) and left untreated, or treated with DTT (10 mM), MB201 (0.3 μM) , or a combination of both (see method). 2 μg of each sample was then left untreated, or treated with subtilisin (1:200 w/w) for 15 min. Samples were then separated by SDS-PAGE and visualized with Coomassie staining. Click here for a detailed method Actin Sedimentation +/- MICAL-1 TreatmentActin (Cat# AKL99) was diluted to 0.2 mg/ml (4.6 μM) or 0.8 mg/ml (18.4 μM) (see method). Samples were then incubated with 2x polymerization buffer or 2x polymerization buffer supplemented with MIC01 + NADPH for 1 h at room temperatire. Samples were spun in an ultracentrifuge at 100,000 g for 1.5 h. Samples were then separated by SDS-PAGE and visualized with Coomassie staining. Click here for a detailed method Actin Polymerization +/- MICAL-1 Treatment.Pyrene labeled-actin (Cat. # AP05) was diluted to 0.1 mg/ml (2.3 μM) (see method). Samples were then incubated with 2x polymerization buffer supplemented with nothing (labeled Control), NADPH alone (labeled +NADPH), MIC01 alone (labeled +MIC01), or the combination of both (labeled +Both). Upon actin polymerization fluorescence was detected with a spectrophotometer (see method below). A.U. = arbitrary units Click here for a detailed method For more information contact: signalseeker@cytoskeleton.comAssociated Products:MOXtrue™ 6xHis MsrB2 Protein (Cat. # MB201)MOXtrue™ MICAL-oxidized Actin (Cat. # MXA95)MOXtrue™ MICAL-oxidized Pyrene Labeled Actin (Cat. # MPAX1)Rabbit Skeletal Muscle Actin (Cat. # AKL95)Pyrene Labeled Rabbit Skeletal Muscle Actin (Cat.# AP05)AboutFor product Datasheets and MSDSs please click on the PDF links below.Certificate of Analysis: Lot001 CitationsFor the most recent publications citing this and other Signal-Seeker™ products, see our Signal-Seeker™ Validation Data Page click hereFaqsVisit our Signal-Seeker™ Tech Tips and FAQs page for technical tips and frequently asked questions regarding this and other Signal-Seeker™ products click here If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.coma
Cytoskeleton，Inc.成立于1993年。自成立以来，我们一直在不断扩大产品范围。
Cytoskeleton，Inc.很高兴为药物筛选，信号转导和细胞骨架研究提供广泛的试剂盒和产品。我们专注于纯化蛋白的生产和易于使用的试剂盒，以研究生化和细胞过程。我们的试剂盒既可用于基础研究或小型筛查的少量样品，也可用于大型筛查的高通量规模
除了我们现有的产品外，我们还提供产品系列中微管，微管蛋白，运动蛋白，小G蛋白效应物，GAP，GEF和其他几种蛋白的药物筛选服务。有关更多信息，请参见我们的药物筛选服务页面。
如果您想从市场上购买到特定产品，请随时与我们联系。我们在这里为您提供帮助。
由于我们的科学家在各自的专业领域都有多年的工作经验，因此我们能够提供高质量的产品。自1993年成立以来，我们以合理的价格生产优质的产品而闻名。
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