Assays


Figure 1. Graph and Table of absorbance Change from Varying Enzyme lab.

Fig. 2. Absorbance change for Run 1 of Varying Substrate lab.

Fig. 3. Absorbance change for Run 2 of Varying Substrate lab.

This week I did enzyme and substrate assays. The enzyme assay turned out well because there were few fluctuations and most of the samples started around 0.5, so there was consistency. I forgot to save data for sample A so it is not on the graph or table, but there was basically no change. Sample F deviated from the general trend so something probably went wrong for that sample. For the substrate assay, I used 60 ul of enzyme. I would have used 70 ul but that would mean I would put in negative 15 ul of nanopure. The first run did not turn out very well because there were more fluctuations and I had to make more enzyme dilution (for samples G and H). There was probably a pippetting error which affected the values for samples G and H. For the second run, more dilution had to be made for sample H. The value for this also did not follow the general trend so there was definitely a pippetting error that affect the enzyme concentration. The absorbance change for run 1 and 2 were about the same (excluding G and H for run 1 and H for run 2) so the results may still be correct despite the pippetting error. I am guessing I would use Sample E or F for the DMSO assay which I will do next week. For the DMSO assay, I will make sure I dilute enough enzyme at the beginning so it limits the chance of a pippetting error. I will probably do inhibition assays the week after Thanksgiving.

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One Response

  1. Thao – yes – I thnk E is what the other guys found is good for CA7. You’ll want to change the x-axis on your graphs when you get to the final report and I would recommend a column graph for the Vary Substrate and Vary DMSO assay graphs but keep the XY graph for the Vary Enzyme.

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