User:Michael F. Nagle/Notebook/Chem 571/2012/11/07

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(Bradford Assay)
(Bradford Assay)
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==Bradford Assay==
==Bradford Assay==
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*A standard curve for UV/Vis was obtained with the following concentrations of BSA:
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*Solutions were prepared with 715μL 1x Bio-Rad Bradford reagant and the following concentrations of BSA:
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*A UV/Vis calibration curve was obtained by analyzing the samples and plotting BSA concentration against absorbance at 595nm.
*The [[User:Michael_F._Nagle/Notebook/Chem_571/2012/11/06|ADA purified yesterday]] was analyzed via UV/Vis and it's peak was compared to the linear regression of BSA's in order to determine concentration.
*The [[User:Michael_F._Nagle/Notebook/Chem_571/2012/11/06|ADA purified yesterday]] was analyzed via UV/Vis and it's peak was compared to the linear regression of BSA's in order to determine concentration.
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==Data==
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[[Image:Bradforassay.png]]
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Fraction #'''
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| align="center" style="background:#f0f0f0;"|'''Absorbance'''
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| align="center" style="background:#f0f0f0;"|'''[ADA] [mg/mL]'''
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| align="center" style="background:#f0f0f0;"|'''molar absorptivity [mL/mg*cm-1]'''
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| align="center" style="background:#f0f0f0;"|'''Molarity (µM)'''
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|-
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| 2||0.93||0.0177||52.4||42.0
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|-
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| 8||0.757||0.0115||65.9||54.4
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|-
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| 9||0.819||0.0137||59.7||65.0
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|-
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|
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|}
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__NOTOC__
__NOTOC__

Revision as of 06:48, 7 December 2012

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Objective

  • Determine concentration of ADA prepared Oct. 23 via Bradford Assay

Bradford Assay

  • Solutions were prepared with 715μL 1x Bio-Rad Bradford reagant and the following concentrations of BSA:
volume BSA (uL) volume H2O (uL [BSA] (mg/mL) [BSA] (M)
03000.00E+00
5250.243.61E-06
10200.152.26E-06
15150.132.01E-06
20100.152.26E-06
2550.243.61E-06
30011.51E-05


  • A UV/Vis calibration curve was obtained by analyzing the samples and plotting BSA concentration against absorbance at 595nm.
  • The ADA purified yesterday was analyzed via UV/Vis and it's peak was compared to the linear regression of BSA's in order to determine concentration.

Data

Image:Bradforassay.png

Fraction # Absorbance [ADA] [mg/mL] molar absorptivity [mL/mg*cm-1] Molarity (µM)
20.930.017752.442.0
80.7570.011565.954.4
90.8190.013759.765.0


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