Based on the data presented above, propose a molecular mechanism by which indibulin is active against cancer cells.
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Based on the data presented above, propose a molecular mechanism by which indibulin is active against cancer cells.
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- You have the following reagents available Rat anti-Keratin 8 antibody specific for mouse Keratin 8, Rabbit polyclonal antibody against mouse keratin 5 and antifade reagent with DAPI to stain DNA. Both primary antibodies work best at 1:500 dilution. A) What additional reagents would you need to perform immunostaining of frozen sections of mouse thymus tissue for Keratin 8 that is expressed in cortical thymic epithelial cells and Keratin 5 that is expressed in mouse medullary thymic epithelial cells? B) Create a staining protocol including all tubes and appropriate controls to stain 4 slides with frozen thymic sections. You will need 200ul of antibody staining solution for each slide.1 of 16 Data from an experiment is shown in the figure below. In the experiment, murine cells were treated with a specific a ligand that activates receptor R. In some cases, the cells were exposed to one of two drugs (X or Y) as well as the ligand or were left untreated (UT). After 30 minutes of treatment, the cells were lysed with a detergent-based buffer to release the soluble membrane, cyosolic and nuclear proteins. Samples from each cell extract were run (in duplicate) by SDS-PAGE (SDS-polyacrylamide gel electrophoresis) in order to separate the proteins by molecular mass (size). The separated proteins were then transferred to a nitrocellulose membrane which was then probed with different antibodies in a western blotting procedure to detect specific phosphorylated proteins or total proteins. If present in the cell extract, these proteins appear as a dark band in the relevant western blot image within the figure. Drug X Drug Y UT Ligand Ligand Ligand Western blotting antibodies…G-protein-coupled receptors on phagocytes link microbe recognition with increased efficiency of intracellular killing. The NBT (Nitro Blue Tetrazolium) test is used to diagnose the genetic disorder Chronic Granulomatous Disease (CGD). To perform this test, peripheral blood cells from the patient are stimulated with bacterial extracts, and then incubated with the NBT compound. Normal neutrophils turn blue in this test due to cleavage of the compound, while patient neutrophils remain uncolored, as shown in Figure below. Name a neutrophil receptor that is likely to be stimulated by the bacterial extract in this assay, and describe how this receptor regulates the activity of the enzyme that cleaves NBT.
- 4 Natalizumab is a therapeutic monoclonal antibody that targets the cell- adhesion molecule a4-integrin and is used as a second-line therapy for severe Crohn's disease. What might be its mechanism of action?类 Protein Specific competitor Mutant/non competitor Add reaction components + +. Probe Antibody Supershift 美美美 Shift Free Probe - Gel separation of complexes Detection of labeled probe Name the experiment shown above and briefly describe how it is set up as well as the role of each component in that setup. What is it used for?refer to the picture explain in detail the type of centrifugation which you will use to seperate imunoglobulin M and immunoglobulin G
- Box one: • Human breast cancer cell line BT474, which overexpresses HER2, in 10 cm Petri dishes• Human HER3 ligand• Lapatinib• Lysis buffer A (containing 1% Triton and 0.1% SDS)• Lysis buffer B (containing no detergent)• Mouse anti-HER3 antibodies• Mouse anti-pY1289 of HER3, that recognises the phospho-tyrosine at position1289 of Her3• Mouse anti-actin antibodies• Horseradish peroxidase (HRP)-conjugated anti mouse IgG antibodies• SDS-PAGE Sample buffer (contains SDS, mercaptoethanol and bromophenolblue)• Molecular weight markers• SDS-PAGE gel• Nitrocellulose membrane• Phosphate buffered saline (PBS)• Blocking buffer (5%w/v BSA in PBS)• X-ray film• Luminol/H2O2 (hydrogen peroxideDescribe how the GPCR and MAPK pathways 'cross-talk' or interact with each other. Give support for your statements. Is cross-talk relevant for drug design? Why or why not?Cell line A was cultured in the absence and presence of KB9520 or PPT. You then lyse the cells and fractionate them, generating membrane, cytosolic and nuclear protein fractions of the whole cell lysate. Western blots were then run using antibodies against ER (alpha) and ER (beta) for each treatment, and include different lanes for the membrane, cytosolic and nuclear protein fractions. Draw the expected results for these western blots and briefly discuss the expected differences.
- According to Research and Development department analysisresults, it was concluded that a two-stage control strategy wasoptimal for antibody production. In the first stage, conditions wereoptimized for cell growth. In the second stage, conditions werechanged to halt cell growth. Please answer the following questions in 1-2 sentences. (a) Why did they choose this two-stage approach? (b) How did they cause the cells to shift from a growth phase to a quiescent (non-growth) phase? (c)Please discuss which factors influence the choice of bioreactor design for optimal antibody production (Hint: The production of antibody which is anchorage-dependentsystems from continuous cell lines can be accomplished usingseveral bioreactor configurations, each of which was designed tosatisfy differing operational, product quality, economic, andproduction scale requirements.)A mutant B cell line is examined by confocal microscopy after incubation with a microbial pathogen recognized by the BCR on these B cells. The B cells have been stained with antibodies to visualize the localization of polymerized actin and microtubules. As a control, wild-type B cells are examined. The results are shown in the figure below, with the numbers indicating the proportion of cells examined that show each pattern of staining. To identify the specific signaling defect in these mutant B cells, a reasonable biochemical assay would be to: Determine if BCR stimulation of mutant B cells produces enhanced binding of the B cell to the microbe Determine whether the mutant B cells have reduced levels of the enzyme Protein kinase C-q Determine whether the mutant B cells are overexpressing the enzyme Vav Determine whether BCR stimulation of mutant B cells promotes exchange of GDP for GTP on cdc42 Determine whether BCR stimulation of mutant B cells produces increased levels of DAG), create a simple flowchart depicting the MC1R pathway. There should be a minimum of five steps in the pathway. Be sure to include reception (protein binding to its ligand), a portion of the transduction pathway (what’s the intracellular reaction? What molecule works intracellularly?), and the cellular response (What’s produced from the cell?).