Acrylamide Solutions and Powders

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Overview

Specifications
Acrylamide Specifications
 
Acrylamide, 99.9% MW: 71.08
conductivity ≤ 2.5 µmho (50% solution)
pH 7.0 ±0.5 (10% solution in a 0.1 M NaCl)
Turbidity ≤ 2.0 NTU (50% sikution)
≤ 1.75 NTU (50% solution in MeOH)
Acrylic acid < 0.002% (determinded indirectly through conductivity and pH measurements. Data available on request.)
 

All acrylamide reagents meet the specifications and have a 1 year guaranteed shelf life.

Premixed Solutions

Premixed Acrylamide:Bis-Acrylamide Solutions

Applications

Acrylamide: Bis-Acrylamide Ratio

Acrylamide/
Bis-Acrylamide Solution, %

Ordering Options

Catalog Number

  • DNA sequencing gels
  • Peptide separation
19:1
(5% cross-linker concentration)
30 500 ml 161-0154
2 x 500 ml 161-0159
40 500 ml 161-0144
2 x 500 ml 161-0145
  • DNA sequencing gels
  • Protein separation
29:1
(3.3% cross-linker concentration)
30 500 ml 161-0156
2 x 500 ml 161-0157
40 500 ml 161-0146
2 x 500 ml 161-0147
  • High molecular weight protein separation
37.5:1
(2.7% cross-linker concentration)
30 500 ml 161-0158
2 x 500 ml 161-0159
40 500 ml 161-0148
2 x 500 ml 161-0149

Premixed Solutions

Concentration,
%

Ordering Options

Catalog Number

Acrylamide 40 500 ml 161-0140
2 x 500 ml 161-0141
Bis-acrylamide 2 500 ml 161-0142
2 x 500 ml 161-0143

Powders
Premixed Acrylamide/Bis-Acrylamide Powder — 99% Pure

Applications

Acrylamide:Bis-Acrylamide Ratio

Ordering
Options

Catalog
Number

  • DNA sequencing gels
  • Peptide separation
19:1
(5% cross-linker concentration)
30 g 161-0120
150 g 161-0123
  • DNA sequencing gels
  • Protein separation
29:1
(3.3% cross-linker concentration)

30 g

161-0121
150 g 161-0124
  • High molecular weight protein separation
37.5:1
(2.7% cross-linker concentration)
30 g 161-0122
150 g 161-0125

Acrylamide Powder

Ordering Options

Catalog Number

100 g 161-0100
500 g 161-0101
1 kg 161-0107
2 kg 161-0103
5 kg 161-0108
Acrylamide/Crosslinker Ratios
Acrylamide/Crosslinker Ratios
Ratio %C Common Applications
19:1 5 DNA sequencing
29:1 3.3 Protein separation
37.5:1 2.6 Protein separation
Crosslinker Application Guide
Crosslinkers
 
  Formal Name Chemical Structure Applications
 
Bis N, N'-methylene-bis-acrylamide General crosslinker in PAGE General crosslinker in PAGE
PDA Piperazine diacrylamide Reduction of silver stain background in SDS-PAGE and 2-D gels, increased resolution, higher gel strength Reduction of silver stain background in SDS-PAGE and 2-D gels, increased resolution, higher gel strength
DATD N, N'-diallylt-tartardiamide Increased pore size of IEF gels where molecular sieving is a problem. Used in scintillation counting: 1,2-diol structure is soluble in periodic acide
 
%T and %C

Pore Size Determination: %T and %C

Polyacrylamide gels are described in terms of two parameters that determine pore size. The total monomer concentration, or %T, is defined as:

%T = ( (grams acrylamide + grams crosslinker) / total volume (ml) ) x 100

The weight percentage of crosslinker, or %C, is defined as:

%C = ( (grams crosslinker) / (grams acrylamide + grams crosslinker) ) x 100

By varying these two parameters, the pore size of the gel can be optimized to give the best separation and resolution for the molecule of interest. For help in determining the best %T and %C for your application, refer to the polyacrylamide gel selection guide section on pages 186–190, which includes examples of migration patterns of proteins on gels of different compositions, or contact Bio-Rad Technical Support.

Gel Types

Gel Types for Specific Applications

SDS-PAGE and native PAGE — Bis-Tris, Tris-acetate, and Tris-HCl gels are available for PAGE applications. SDS-PAGE is the most common protein electrophoretic application. It separates complex protein mixtures by size, allowing estimation of the molecular weight of sample proteins. Bio-Rad’s Tris-HCl gels are made without SDS. By omitting SDS from the running buffer,the gels can be used to run proteins under nondenaturing conditions for subsequent analysis of their native conformation and activity. Native analysis does not provide accurate molecular weight information. Gels for SDS-PAGE and native PAGE protein separations have traditionally been formulated with Tris-HCl. The neutral-pH formulations of Bis-Tris and Tris-acetate gels significantly delay acrylamide hydrolysis and give these gels longer shelf life (12 months for Bis-Tris gels, 8 months for Tris-acetate gels) than conventional Laemmli Tris-HCl gels. By using optimized combinations of Criterion XT neutral-pH gel buffers and running buffers, all sizes of proteins can be separated into tight, well-resolved bands on a range of only two or three acrylamide percentages. Criterion XT gels can also be used for SDS-PAGE or native PAGE applications.

Small protein and polypeptide analysis — Tris-Tricine gels are ideal for separation of peptides and small proteins with molecular weight <10,000. Superior resolution is achieved by slowing the migration rate of the peptide-SDS complexes. This helps achieve separation from the faster-moving SDS micelles that interfere with peptide resolution in Tris-glycine buffer systems.

IEF — IEF gels separate proteins based on their net charge rather than molecular weight. IEF gels are cast with carrier ampholytes to create a pH gradient within the gel. Proteins migrate to their pI, the pH at which their net charge is zero. IEF gels contain no denaturing agents, allowing one-dimensional separation under native conditions.

Protease detection — Zymogram gels contain either gelatin or casein, which are substrates for various proteases. Samples are run under denaturing but nonreducing conditions, then allowed to renature and consume the substrate. Samples with proteolytic activity can be visualized as clear bands against a blue background after Coomassie Blue R-250 staining.

dsDNA separation — TBE gels are suitable for electrophoresis of nucleic acids from 50 to 2,000 base pairs; they are ideal for analysis of the purity of PCR products, standard dsDNA analysis, and RNase protection assays.

ssDNA and RNA separation — TBE-urea gels maintain denaturing conditions for analysis of single-stranded DNA and RNA. Nucleic acids between 60 and 200 bases are resolved as sharp, distinct bands. Applications include oligonucleotide purity analysis, RNase protection assays, and northern blotting.

Acrylamide Percentage (%T)

Proper acrylamide concentration is critical to the success of electrophoretic separation. Acrylamide concentrations that are too high can lead to exclusion of high molecular weight molecules from the gel; concentrations that are too low can decrease sieving. Use the protein and DNA migration charts (pages 188 190) to select a gel with a %T that will provide optimal resolution of your sample.

Single-percentage gels — Choose a single-percentage gel when your sample has a limited size range of molecules and your goal is to separate a single band from neighboring bands. In general, single-percentage gels will produce the greatest separation between bands with similar molecular weights. Single-percentage Tris-HCl, Bis-Tris, Tris-acetate, Tris-Tricine, and zymogram gels are cast with a 4% stacking gel to further sharpen protein bands before they enter the resolving gel.

Gradient gels — Choose a linear gradient gel if your sample contains a wide range of molecular weights. These gels allow both high and low molecular weight bands to be visualized on the same gel.

Crosslinker Percentage (%C)

Crosslinker percentage is the weight percentage of crosslinker (bis-acrylamide). Along with the total monomer concentration or %T (acrylamide), %C determines the pore size of the gel. Crosslinker percentage can be adjusted to optimize pore size in order to deliver the best separation and resolution for the molecule of interest. Bio-Rad precast gels have a %C optimized for the bestseparation of molecules for their applications. Refer to the table below for the crosslinker percentage in Bio-Rad precast gels.

Approximate number of 12% mini gels that can be run
Product Description Number of 12% Mini Gels
500 ml 40% Acrylamide/Bis Solution 300
500 ml 30% Acrylamide/Bis Solution 225
150 g Premixed Acrylamide/Bis Powder 225
30 g Premixed Acrylamide/Bis Powder 45
100 g Acrylamide Powder 150

Acrylamide solutions and powders are used to generate polyacrylamide, a chemically inert, electrically neutral, and hydrophilic gel matrix used for separating biomolecules by polyacrylamide gel electrophoresis (PAGE). The reliability of electrophoresis results depends on the reproducibility of this reaction, which is dependent on several factors.

The polymerization process requires bis-acrylamide as a cross-linking agent. Adjusting the concentration and ratio of acrylamide and bis-acrylamide creates gel matrices with a range of pore sizes that allow the electrophoretic separation of nucleic acids, peptides, and proteins — molecules ranging in molecular weight from 2 to 5,000 Da.

The purity of acrylamide reagents used in the polymerization process determines the uniformity of the gel matrix. Bio-Rad offers a range of 99.9% pure reagents to suit your DNA sequencing (6, 8, 12, 20% acrylamide), and/or peptide and protein electrophoresis applications. Our choices of pure acrylamide and/or bis-acrylamide premixed solutions or powders include:

Convenient ordering options are available to meet your acrylamide handcasting gel demands. Bio-Rad’s stock solutions and powders can be used to prepare

  • Single-percentage gels (7.5–20%) — for separating bands that are close in molecular weight
  • Gradient gels (4–15% and 10–20%) — for separating samples containing a broad range of molecular weights
  • Broad gradient gels (4–20% and 8–16%) — for evaluating new or unknown samples

More Information

Our accessories and reagents for handcasting multiple gels include polyacrylamide gel reagents, premade buffers and reagents, multicasting chambers for mini, midi, large gels, and gradient formers.

View our wide selection of mini and midi precast gels, electrophoresis equipment available in mini and midi formats, and protein standards for your specific protein separation applications.

Learn more about casting polyacrylamide gels, protein electrophoresis techniques, and types of protein electrophoresis (PAGE).

30% Acrylamide/Bis Solution, 19:1

161-0154
500 ml, 30% acrylamide and bis-acrylamide solution, 19:1

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30% Acrylamide/Bis Solution, 29:1

161-0156
500 ml, 30% acrylamide and bis-acrylamide solution, 29:1

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30% Acrylamide/Bis Solution, 37.5:1

161-0158
500 ml, 30% acrylamide and bis-acrylamide solution, 37.5:1

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40% Acrylamide/Bis Solution, 19:1

161-0144
500 ml, 40% acrylamide and bis-acrylamide solution, 19:1

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40% Acrylamide/Bis Solution, 29:1

161-0146
500 ml, 40% acrylamide and bis-acrylamide solution, 29:1

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40% Acrylamide/Bis Solution, 37.5:1

161-0148
500 ml, 40% acrylamide and bis-acrylamide solution, 37.5:1

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40% Acrylamide Solution

161-0140
500 ml, 40% acrylamide solution

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2% Bis Solution

161-0142
500 ml, 2% bis-acrylamide solution

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30% Acrylamide/Bis Solution, 19:1

161-0155
2 x 500 ml, 30% acrylamide and bis-acrylamide solution, 19:1

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30% Acrylamide/Bis Solution, 29:1

161-0157
2 x 500 ml, 30% acrylamide and bis-acrylamide solution, 29:1

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30% Acrylamide/Bis Solution, 37.5:1

161-0159
2 x 500 ml, 30% acrylamide and bis-acrylamide solution, 37.5:1

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40% Acrylamide/Bis Solution, 19:1

161-0145
2 x 500 ml, 40% acrylamide and bis-acrylamide solution, 19:1

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40% Acrylamide/Bis Solution, 29:1

161-0147
2 x 500 ml, 40% acrylamide and bis-acrylamide solution, 29:1

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40% Acrylamide/Bis Solution, 37.5:1

161-0149
2 x 500 ml, 40% acrylamide and bis-acrylamide solution, 37.5:1

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40% Acrylamide Solution

161-0141
2 x 500 ml, 40% acrylamide solution

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2% Bis Solution

161-0143
2 x 500 ml, 2% bis-acrylamide solution

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Acrylamide

161-0100
100 g, 99.9% pure acrylamide powder

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Acrylamide

161-0101
500 g, 99.9% pure acrylamide powder

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Acrylamide

161-0107
1 kg, 99.9% pure acrylamide powder

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Acrylamide

161-0103
2 kg, 99.9% pure acrylamide powder

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Acrylamide

161-0108
5 kg, 99.9% pure acrylamide powder

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Acrylamide/Bis Powder

161-0120
30 g, 19:1 mixture of 99.9% pure acrylamide and bis-acrylamide powder

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Acrylamide/Bis Powder

161-0121
30 g, 29:1 mixture of 99.9% pure acrylamide and bis-acrylamide powder

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Acrylamide/Bis Powder

161-0122
30 g, 37.5:1 mixture of 99.9% pure acrylamide and bis-acrylamide powder

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Acrylamide/Bis Powder

161-0123
150 g, 19:1 mixture of 99.9% pure acrylamide and bis-acrylamide powder

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Acrylamide/Bis Powder

161-0124
150 g, 29:1 mixture of 99.9% pure acrylamide and bis-acrylamide powder

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Acrylamide/Bis Powder

161-0125
150 g, 37.5:1 mixture of 99.9% pure acrylamide and bis-acrylamide powder

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Number Description Options
LIT306 Instruction Manual, Premixed, Preweighed Acrylamide:Bis, Rev C Click to download
1156 Acrylamide Polymerization — A Practical Approach Click to download
1866 Acrylamide Brochure Click to download
LIT492 Instruction Manual, Acrylamide and bis-Acrylamide Solutions Click to download [ Add to Cart (Free) ]
2317 Ready-to-Run Buffers and Solutions Brochure, Rev F Click to download [ Add to Cart (Free) ]