User:Benjamin Friedel/Notebook/CHEM 471/2015/10/27: Difference between revisions
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## Through M1V1= M2V2 the correct volume of protease needed for a final incubation tube concentration of 1uM and final tube volume of 1mL was determined | ## Through M1V1= M2V2 the correct volume of protease needed for a final incubation tube concentration of 1uM and final tube volume of 1mL was determined | ||
### V1= 4.59uL | ### V1= 4.59uL | ||
# Each sample and blank tube for incubation requires a total volume of 1mL, thus 1mLtotal - 4.59uL protease = 995.41uL | # Each sample and blank tube for incubation requires a total volume of 1mL, thus 1mLtotal - 4.59uL protease = 995.41uL Tris buffer. | ||
'''Incubation Tube Preparation (Samples and Blanks)''' | '''Incubation Tube Preparation (Samples and Blanks)''' | ||
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### UV Absorbance was then immediately measured from 400-800nm | ### UV Absorbance was then immediately measured from 400-800nm | ||
# Note that a spectra with the same parameters was taken for a sample with the same makeup as shown above, with the replacement of the incubated sample with Tris buffer for a bradford blank. | # Note that a spectra with the same parameters was taken for a sample with the same makeup as shown above, with the replacement of the incubated sample with Tris buffer for a bradford blank. | ||
==Data== | ==Data== |
Revision as of 12:55, 22 November 2015
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ObjectiveThe objective for today is to measure the UV absorbance of lysozyme AuNP fibers over various incubation times with alpha chymotrypsin using a bradford assay. ProtocolProtease Preparation
Incubation Tube Preparation (Samples and Blanks)
DataFigure 1
Figure 2 above shows the absorbance data of the AuNP samples over alpha-chymotrypsin incubation corrected for blanks. Blank correction was performed by subtracting the values of the Bradford blank and isosbestic point corrected blank data from the Bradford blank and isosbestic point corrected sample (AuNP fiber tube) data. The increase in absorbance as incubation time increases suggests that as alpha-chymotrypsin digests AuNP fibers samples, peptides and small fiber particles are released into solution. |