Exp. 2  Protein Fractionation and Quantification 

I.                          Object

Please use two sentences to describe the purpose of this experiment. 

II.                        Methods

Please briefly describe the processes involved.

1.    Fractionation:

·         Describe how you obtain the 100% protein sample from egg white).

2.    Quantification:

·         Preparation of BSA solution from stock.

·         Bradford assay 

III.                     Results

1.    Your OD₅₉₅ data

2.    Construct the standard curve by plotting OD₅₉₅ of the standards versus protein concentration of the known.

3.    Calculate the protein concentration of the unknowns (the original crude albumin) from the standard curve and the dilution factors if necessary. 

IV.                    Question and Discussion

1.    Discuss the mechanisms for salt and organic solvent precipitation and the advantage and disadvantage of both methods in protein purification.

2.    Find another commonly used assay for determining protein concentration and briefly describe its basic principle.  Compare the advantage and disadvantage with the Bradford assay we used in class.

3.    Other things you want to discuss.

V.                      Reference

1.    Please cite the references if you use any, including the web resources.

VI.      Common mistakes

1.    No. of significant digits below the decimal.

2.    Graphing in Excel:  X-Y 散佈圖 vs. 折線圖

利用 Excel 作圖要選擇正確的作圖方式:

Data:

   

用同樣的數據 下圖是一個選錯作圖方式的例子:

(在此橫軸上, 每點間的距離是一樣的....,  雖然依然可以找到一條直線, 但卻是非常不一樣的結果!!)

                                                                            

3.    Title of the graph, legends, and label of the X-, Y-axis, and units.

4.    Unknown not “unknow”.

5.    BSA (bovine serum albumin) not “ABS”, “BAS”.

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上次更新日期： 2005年08月11日。