Question
Asked 18th Mar, 2015

What is the difference between acrylamide and bisacrylamide?

I am a bit confused by the difference between acrylamide and bisacrylamide powder. Are both powders the same or not? Can I used acrylamide only for my SDS-PAGE instead of using bisacrylamide?

Most recent answer

20th Dec, 2021
Cássio Cassiano
Universidade Federal São João del-Rei
A agree with all comments.
Both reagents are necessary for your gel.
At this moment I work with 2 proteins (25 kDa and 27 kDa), and I use a 30% Acrylamide and 0,8% Bis Acrylamide solution (30 g Acrylamide + 0,8 gramas Bis).
I recommend: Filter the solution and always using a fresh solution (3 months are good to use).
Check this article:
I use this protocol in my lab.

Popular Answers (1)

18th Mar, 2015
Prem Prakash Das
National Research Council Canada
Acrylamide and Bisacrylamide, both are very important for proper polymerization. Acrylamide forms linear polymers whereas Bisacrylamide cross links these linear polymer. More numbers of cross links means smaller pore size. So, ratio of Acrylamide and Bisacrylamide determines pore size.
24 Recommendations

All Answers (10)

18th Mar, 2015
Prem Prakash Das
National Research Council Canada
Acrylamide and Bisacrylamide, both are very important for proper polymerization. Acrylamide forms linear polymers whereas Bisacrylamide cross links these linear polymer. More numbers of cross links means smaller pore size. So, ratio of Acrylamide and Bisacrylamide determines pore size.
24 Recommendations
18th Mar, 2015
Ron Reade
Agriculture and Agri-Food Canada
Hi Rasdin,
Prem is correct, you need both (points for you Prem). Just to help out a little more, we generally use a 30% Bis-Acrylamide solution for our standard SDS-PAGE. so 87.6g Acrylamide + 2.4g Bis-Acrylamide in 300mls final water. Just remember that acrylamide is a cumulative neurotoxin and is very dangerous to inhale the powder.
Cheers
Ron 
12 Recommendations
19th Mar, 2015
Mark K Chee
Martin Methodist College
Here is an excellent article from National Diagnostics on this topic: https://nationaldiagnostics.com/electrophoresis/article/polyacrylamide-matrix
"For discussions of the composition of polyacrylamide gels, a standard nomenclature has been widely adopted. In this nomenclature, T represents the total percentage concentration (w/v) of monomer (acrylamide plus crosslinker) in the gel. The term C refers to the percentage of the total monomer represented by the crosslinker. For example, an 8%, 19:1 (acrylamide/bisacrylamide) gel would have a T value of 8% and a C value of 5%."
7 Recommendations
19th Mar, 2015
Mark K Chee
Martin Methodist College
Rasdin, do you intend to analyze proteins or nucleic acids on your polyacrylamide gel?
19th Mar, 2015
Rasdin Ridwan
Universiti Teknologi MARA
Tq Prem for the answer and Ron for the explanation...really appreciated that.
Mark, i intend to analyzes p53 and bax protein...
19th Mar, 2015
Mark K Chee
Martin Methodist College
I'm assuming that you will also be analyzing a loading control protein within the size range of your two proteins of interest. Bax should be about 20 kDa, am I correct? You can try  starting with a 12.5%, 37.5:1 (acrylamide/bisacrylamide) gel in that case to get good separation in that molecular weight range (20-50 kDa). I hope that helps.
2 Recommendations
1st Apr, 2015
Mark K Chee
Martin Methodist College
Relevant paragraphs from the National Diagnostics article:
Control of the pore size of a polyacrylamide gel is accomplished by changing the T and C values. With increasing T, the pore size decreases in a nearly linear relationship. Higher percentage gels (higher T), with smaller pores, are used to separate smaller molecules. The relationship of C to pore size is more complex. Generally, the minimum pore size occurs when C is about 5% (a 19:1 gel). Decreasing C results in a more open pore structure because there are fewer crosslinker molecules. Increasing C beyond 5% also increases the pore size. This appears to be because of nonhomogeneous bundling of strands in the gel.
Researchers have settled on C values of 5.0% (19:1 acrylamide/bis) for most forms of denaturing DNA and RNA electrophoresis and 3.3% (29:1) for most native DNA and RNA gels. For SDS-PAGE electrophoresis of proteins, the standard C value that has been adopted is 2.6% (37.5:1)."
7 Recommendations
18th Jul, 2016
Tayyeb Allahverdi Pourfallah
Mazandaran University of Medical Sciences
By the way, acrylamide is very toxic.
3rd Mar, 2020
Seob YONG Jung
Ajou University

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