Title

Quantitative Application for SDS-PAGE in an Undergraduate Biochemistry Lab

Presenter Information

Brandon Petersen
Sarah Printz
John Carter

Document Type

Oral Presentation

Location

SURC Ballroom A

Start Date

17-5-2012

End Date

17-5-2012

Abstract

The most common technique for separating and visualizing proteins in their native state is polyacrylamide gel electrophoresis (PAGE), while the incorporation of sodium dodecyl sulfate (SDS-PAGE) is often used to analyze denatured proteins. SDS-PAGE is frequently utilized in undergraduate teaching labs to analyze the composition of a sample of protein(s) and/or to determine the molecular weight of a particular protein. However, quantitatively determining the concentration of a particular protein within a mixture has generally required the implementation of expensive imaging and gel analysis software. Here, we describe an inexpensive protocol to accurately quantitate the concentration of a single protein from a mixture of serum proteins using this common technique. Briefly, this exercise uses SDS-PAGE to separate the protein components of human serum, along with a standard curve of bovine serum albumin (BSA). The resulting gel is then dried between sheets of cellophane and scanned with a common flatbed scanner. Free software from the National Institute of Health is then used to quantitate the amount of albumin in the sample of human serum. This protocol produces consistently reliable results at a dramatically reduced cost when compared to standard protocols that require a laser densitometer and/or expensive imaging equipment and analysis software.

Poster Number

29

Faculty Mentor(s)

Timothy Sorey, Todd Kroll

Additional Mentoring Department

Chemistry

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May 17th, 8:30 AM May 17th, 11:00 AM

Quantitative Application for SDS-PAGE in an Undergraduate Biochemistry Lab

SURC Ballroom A

The most common technique for separating and visualizing proteins in their native state is polyacrylamide gel electrophoresis (PAGE), while the incorporation of sodium dodecyl sulfate (SDS-PAGE) is often used to analyze denatured proteins. SDS-PAGE is frequently utilized in undergraduate teaching labs to analyze the composition of a sample of protein(s) and/or to determine the molecular weight of a particular protein. However, quantitatively determining the concentration of a particular protein within a mixture has generally required the implementation of expensive imaging and gel analysis software. Here, we describe an inexpensive protocol to accurately quantitate the concentration of a single protein from a mixture of serum proteins using this common technique. Briefly, this exercise uses SDS-PAGE to separate the protein components of human serum, along with a standard curve of bovine serum albumin (BSA). The resulting gel is then dried between sheets of cellophane and scanned with a common flatbed scanner. Free software from the National Institute of Health is then used to quantitate the amount of albumin in the sample of human serum. This protocol produces consistently reliable results at a dramatically reduced cost when compared to standard protocols that require a laser densitometer and/or expensive imaging equipment and analysis software.