Quantitative Application for SDS-PAGE in an Undergraduate Biochemistry Lab
Document Type
Oral Presentation
Campus where you would like to present
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.
Recommended Citation
Petersen, Brandon; Printz, Sarah; and Carter, John, "Quantitative Application for SDS-PAGE in an Undergraduate Biochemistry Lab" (2012). Symposium Of University Research and Creative Expression (SOURCE). 101.
https://digitalcommons.cwu.edu/source/2012/posters/101
Poster Number
29
Additional Mentoring Department
Chemistry
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.
Faculty Mentor(s)
Timothy Sorey, Todd Kroll