1. Use the following Absorbance Data to generate a Bradford Assay Standard Curve.Protein ConcentrationAbsorbance at 595nm(mg/mL)0.05Chart Title0.00920.10.020.20.040.30.1120.40.1670.50.19800.60.2436Absorbance at 595nm0.34y = 0.4499x - 0.0254R? = 0.98240.250.20.150.10.050.10.20,30.40,50.60.7 2. Use the linear regression equation and the following Abs 595 data and dilution factors forcalculating your protein concentrations:SAMPLEABS 595DILUTIONCalculatedFACTOR[protein](mg/mL)CRUDE0.114103.098FLOW THRU0.1195PRE DESALTING0.1763POST DESALTING0.15733. Using the protein concentrations that you calculated, determine the volume in ul of each samplthat would be needed to load 25ug of that sample on an SDS-PAGE. See example in the first row ofthe table.SAMPLEVolume ofVol of 4xVolume ofFinal totalvolume (ul)sample for25ug (uL)gelloadingbufferTris forsampleCRUDE8251750FLOW THRU50PRE50DESALTINGPOST50DESALTING

Answered: Image /qna-images/answer/6a90c364-2dc0-42b5-af44-78e309c447a0.jpg

2. Usé thè linear regression équation ànd the fóllowing Abs 595 dată and dilution factors för calculating your protein concentrations: SAMPLE ABS 595 DILUTION Calculated FACTOR [protein] (mg/mL) 3.098 CRUDE 0.114 0.119 0.176 10 FLOW THRU 5 PRE DESALTING 3 POST DESALTING 0.157 3

2. Usé thè linear regression équation ànd the fóllowing Abs 595 dată and dilution factors för calculating your protein concentrations: SAMPLE ABS 595 DILUTION Calculated FACTOR [protein] (mg/mL) 3.098 CRUDE 0.114 0.119 0.176 10 FLOW THRU 5 PRE DESALTING 3 POST DESALTING 0.157 3

Appl Of Ms Excel In Analytical Chemistry

2nd Edition

ISBN:9781285686691

Author:Crouch

Publisher:Crouch

Chapter12: Spectrochemical Methods

Section: Chapter Questions

Problem 4P

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Concept explainers

Mass Spectrometry

This process is an instrumental method in which a sample is changed to fast moving positive ions by electron bombardment. The charged particles vary due to their different mass to charge ratio and are estimated with the help of this difference. It is also called mass spectroscopy.

Question

1. Use the following Absorbance Data to generate a Bradford Assay Standard Curve.
Protein Concentration
Absorbance at 595nm
(mg/mL)
0.05
Chart Title
0.0092
0.1
0.02
0.2
0.04
0.3
0.112
0.4
0.167
0.5
0.1980
0.6
0.2436
Absorbance at 595nm
0.3
4y = 0.4499x - 0.0254
R? = 0.9824
0.25
0.2
0.15
0.1
0.05
0.1
0.2
0,3
0.4
0,5
0.6
0.7

2. Use the linear regression equation and the following Abs 595 data and dilution factors for
calculating your protein concentrations:
SAMPLE
ABS 595
DILUTION
Calculated
FACTOR
[protein]
(mg/mL)
CRUDE
0.114
10
3.098
FLOW THRU
0.119
5
PRE DESALTING
0.176
3
POST DESALTING
0.157
3
3. Using the protein concentrations that you calculated, determine the volume in ul of each sampl
that would be needed to load 25ug of that sample on an SDS-PAGE. See example in the first row of
the table.
SAMPLE
Volume of
Vol of 4x
Volume of
Final total
volume (ul)
sample for
25ug (uL)
gel
loading
buffer
Tris for
sample
CRUDE
8
25
17
50
FLOW THRU
50
PRE
50
DESALTING
POST
50
DESALTING

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