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I'm a pharmacy student and I can try to explain how they do to find new drugs. In fact, there are many diferent ways to do so, it depends of what kind of new drug you are searching for. Sometimes you're working on a disease which the pathological molecular mechanism of action are well-known and where many drugs are already effective. In this case, the '...


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An assay may be thought of as a trial or test that is designed to quantitatively determine the amount of a substance in a sample. Thus a biochemist might use a protein assay to quantitatively determine the amount of protein in a sample, or a pharmacologist might assay a sample to quantitatively determine the amount of drug present, or an enzymologist ...


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I want to add something on top of Chris answer. The production of an antibody it is usually a quite slow (and expensive) process, an alternative that worth to consider is phage display (https://en.wikipedia.org/wiki/Phage_display). Once you find the phage that effectively bind your protein of interest, you can use it instead of an antibody in what is called ...


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I think the most promising routes use antibodies. You could either develop an ELISA or do western blot analysis of plant material - both need a good and specific antibody. To generate these, the protein of interest (or at least parts of it) are injected into animals (typically mice or rabbit) and then antibodies are pourified from the blood of these animals. ...


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Galvinoxyl is a free radical, which means it has an unpaired electron that can be detected with EPR. Antioxidants reduce the radical, which is then not detectable in the EPR anymore as it doesn't have an unpaired electron. What you need the EPR for in the assay is to determine how much of the galvinoxyl is still a radical. Paramagnetic molecules like ...


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Some ATPases can work with MANT-ATP or similar fluorescent ATP analogs that change their fluorescence properties upon protein binding as well as hydrolysis of their phosphate group. This has been used frequently (see here or here) to study enzyme kinetics on fast timescales.


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I have not used the assay myself, but Bhuiya and Liu "A cost-effective colorimetric assay for phenolic O-methyltransferases and characterization of caffeate 3-O-methyltransferases from Populus trichocarpa." Anal Biochem. 2009 384(1):151-8 [link] seems to have a relatively straightforward protocol. Roughly, you make a 0.4% (w/v) solution of Gibbs reagent in ...


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In my experience, reuse of NADPH stock solutions maintained at pH 7.0 or above is perfectly acceptable. Taking an absorbance spectrum every half-hour or so will probably show that definitively. However, NADPH is VERY unstable in acid, leading to a 'bleaching' of the absorbance at 340nm. Try measuring a rate at pH 6 or below, 340nm, with 100 micromolar ...


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Using different wavelength for reference give some advantages. You can measure a baseline of each well. All well might not have the same baseline because each well might not be the exactly same as others. In addition, you could make scratches on it, you might touch the bottom, or you get some dirt from your bench. And this can be corrected by A540 in your ...


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So, your blank is everything but the enzyme? That is, substrate and DNSA with 100 uL water? If yes, measure, as negative controls, substrate alone and DNSA alone. If one of them is also highly absorbing light, it may be contaminated, and a new vial should be ordered. If both are highly absorbing, it may be the water you are using, or the spectrophotometer. ...


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Background: There are a lot of recipes to extract protein from human tissues out there, but all of them boil down to one thing; preserving the tissue proteins as much as possible while obtaining a reasonable yield for downstream applications, using extraction buffer, extra techniques, and by adding protease inhibitors ELISA is one of the those ...


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It's hypothesized that planar cell polarity is many cases functionally associated with asymmetry of (primary) cilia (Axelrod (2008)). So, the phenomenon of primary cilium migration could be not that infrequent. One example is the development of ependymal cells in brain ventricules: They are multiciliated and the cilia are motile, but ciliogenesis starts from ...


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Perhaps you're looking for these hanging inserts, for example: "Millicells"


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Apparently poly(ethylene oxide) is a biologically interesting polymer, with minimal protein interactions. The apparently means that I haven't tried it myself, but my polymer teacher told us so. http://www.sigmaaldrich.com/materials-science/material-science-products.html?TablePage=20204110 http://en.wikipedia.org/wiki/Polyethylene_glycol Nomenclature ...



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