Electrophoresis; Separation of Proteins
Purpose: To create an electrophoresis gel in order to further separate Cytochrome C, Myoglobin, Albumin and Hemoglobin(proteins) based off of their net charges.
Materials:
• Rabbit hemoglobin
• Serum albumin(cow)
• Myoglobin
• Cytochrome C
• Microtubes
• Tape
• Micropipets
• Agarose gel
• Buffer
• Gel chamber
• Test tubes
• Test tube holders
• Gloves
• Macropipets
Properties of Proteins:
Protein Color Isoelectric Point Net Change at pH 8.6
Cytochrome C Orange 10.2 Positive
Myoglobin Brown-red 7.2 Negative
Hemoglobin Red 6.8 Negative
Serum Albumin Blue 4.8 Very negative
Introduction: Electrophoresis, in general, is a commonly utilized technique for separating charged macromolecules depending on their size. It is also used to analyze both DNA and RNA. The process of electrophoresis includes using an electric current to differentiate molecules based on size in a sponge-like matrix. The smaller of the molecules moves more easily through the gel than do the large molecules. The matrix used in gel electrophoresis is agarose. Agarose is actually a chain of sugar molecules derived from seaweed and in its pure form, is not soluble in water at room temperature. However, it does dissolve in boiling water. As …show more content…
10 microliters of each of the four proteins were to be placed into the sample wells with a couple of sample wells left in between each of them in order to more easily compare the end results and avoid collision of substances. After making sure that each of the gels were transferred to the correct electrophoretic cell, the chamber was to be electrophoresed for 8 minutes before it was to be turned off. The location of each of the four proteins was to be recorded compared to the insertion point. Electrophoresis was to be continued if bromophenol blue (within the serum albumin) sample had not yet moved close to the positive end of the
Purpose: To create an electrophoresis gel in order to further separate Cytochrome C, Myoglobin, Albumin and Hemoglobin(proteins) based off of their net charges.
Materials:
• Rabbit hemoglobin
• Serum albumin(cow)
• Myoglobin
• Cytochrome C
• Microtubes
• Tape
• Micropipets
• Agarose gel
• Buffer
• Gel chamber
• Test tubes
• Test tube holders
• Gloves
• Macropipets
Properties of Proteins:
Protein Color Isoelectric Point Net Change at pH 8.6
Cytochrome C Orange 10.2 Positive
Myoglobin Brown-red 7.2 Negative
Hemoglobin Red 6.8 Negative
Serum Albumin Blue 4.8 Very negative
Introduction: Electrophoresis, in general, is a commonly utilized technique for separating charged macromolecules depending on their size. It is also used to analyze both DNA and RNA. The process of electrophoresis includes using an electric current to differentiate molecules based on size in a sponge-like matrix. The smaller of the molecules moves more easily through the gel than do the large molecules. The matrix used in gel electrophoresis is agarose. Agarose is actually a chain of sugar molecules derived from seaweed and in its pure form, is not soluble in water at room temperature. However, it does dissolve in boiling water. As …show more content…
10 microliters of each of the four proteins were to be placed into the sample wells with a couple of sample wells left in between each of them in order to more easily compare the end results and avoid collision of substances. After making sure that each of the gels were transferred to the correct electrophoretic cell, the chamber was to be electrophoresed for 8 minutes before it was to be turned off. The location of each of the four proteins was to be recorded compared to the insertion point. Electrophoresis was to be continued if bromophenol blue (within the serum albumin) sample had not yet moved close to the positive end of the