Assignment: Cell Fractionation Worksheet

Assignment: Cell Fractionation Worksheet

Assignment: Cell Fractionation Worksheet

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All questions are listed. All data is listed. This was a pre-preformed lab.

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Worksheet for cell Fractionation, Chapter 8

Introduction

Cellular fractionation is a technique that enables researchers to isolate cellular organelles. Once isolated, the presence and functions of these organelles can be studied biochemically.

1. Characterization of Mitochondria by Succinic Dehydrogenase Assay

The presence of mitochondria is measured by the ability of the fraction to metabolize succinate and reduce DCPIP (blue) to DCPIPH2 (clear).

Procedure: Described in Powerpoint and in Teams recording

Table1.

1. Which sample (fraction) contains higher concentration of mitochondria? Explain why. (4 points)
2. Characterization of Mitochondria by Differential Spectral Analysis

Procedure: Described in Powerpoint and in Teams recording

Results:

Table 2: absorbance values of scan of fractions 1 and 2.

1. Graph the data above on the same axis with Absorbance on Y-axis ( points) and wavelength on the X-axis. (4 points)

2. What is the role of CaCl2 used in the spectral analysis? (2 points)

Note: Since the absorbance is directly proportional to the concentration, we will also these abs as concentrations in mM.

3. Use the graph and determine the concentrations of CytC; CytC reductase, and CytC oxidase and complete table 3 (6 points)

Table 3. Abs values (or mM)

4. Which form of cytochrome C is more predominant in fraction 2? (2 points)
5. Attended the lab? _____________ (2 points)

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