Protein Electrophoresis Buffers and Reagents bottle

Finding the right protein gel electrophoresis system is critical to getting consistent and accurate results you need for your research. Electrophoresis buffers and reagents are important components of the protein electrophoresis system. We offer a range of SDS-PAGE buffers, native buffers and reagents for gel casting, sample preparation, running, and transferring gels. Find the recommended electrophoresis buffers and reagents for each gel system below.

Protein gels  Chamber systems

SDS sample buffers or loading buffers

Protein samples prepared for SDS-PAGE analysis are denatured by heating in the presence of a sample buffer containing 1–2% SDS or LDS with or without a reducing agent such as 20 mM DTT, 2-mercaptoethanol (BME) or TCEP. Sample buffers or loading buffers also contain glycerol so that they are heavier than water and sink neatly to the bottom of the buffer-submerged well when added to a gel. In addition, a negatively charged, low-molecular weight dye is also included in the sample buffer that will migrate at the buffer-front, enabling one to monitor the progress of electrophoresis. The most common tracking dyes for sample loading buffers are bromophenol blue, phenol red, and Coomassie blue. The table below summaries common sample buffers used in the different gel buffer systems.

Gel system Denaturing sample buffer (final concentrations) Sample preparation
Tris-glycine Tris-glycine SDS sample buffer: Tris HCl (63 mM), glycerol (10%), SDS (2%), bromophenol blue (0.0025%), pH 6.8 Heat samples at 85°C for 2–5 minutes with sample buffer
Bis-Tris LDS sample buffer: Tris base (141 mM), Tris HCl (106 mM), LDS (2%), EDTA (0.51 mM), SERVA Blue G-250 (0.22 mM), phenol red (0.175 mM), pH 8.5 Heat samples at 70°C for 10 minutes with sample buffer
Tris-Acetate LDS sample buffer: Tris base (141 mM), Tris HCl (106 mM), LDS (2%), EDTA (0.51 mM), SERVA Blue G-250 (0.22 mM), phenol red (0.175 mM), pH 8.5 Heat samples at 70°C for 10 minutes with sample buffer
Tris-Tricine Tricine SDS sample buffer: Tris HCl (450 mM), glycerol (12%), SDS (4%), Coomassie Blue G (0.00075%), phenol red (0.0025%), pH 8.45 Heat samples at 85°C for 2–5 minutes with sample buffer
Zymogram Tris-glycine SDS: Tris HCl (63 mM), glycerol (10%), SDS (2%), bromophenol blue (0.0025%), pH 6.8 Do not heat samples, load immediately onto gel after adding to sample

For fluorescence applications, we recommend that you use sample buffers that do not contain dyes that interfere in fluorescent channels (e.g., bromophenol blue). Fluorescent Compatible Sample Buffer does not contain bromophenol blue or other interfering dyes and can be used with reducing or nonreducing SDS-PAGE applications.

Find SDS sample buffer recipes in gel-specific product manuals in the documents tab.

Native sample buffers

Protein samples prepared for native PAGE applications are prepared in sample buffers or loading buffers that do not contain detergents to maintain the native state of the proteins. The loading buffers contain glycerol so that they are heavier than water and sink neatly to the bottom of the buffer-submerged well when added to a gel. In addition, a negatively charged, low-molecular weight dye is also included in the sample buffer that will migrate at the buffer-front, enabling one to monitor the progress of electrophoresis.

When preparing samples for native applications, protein samples should not be heated and loaded immediately onto the gel after mixing with the sample buffer.

Gel system Native sample buffer (final concentrations)
Tris-glycine Native sample buffer: Tris HCl (100 mM), glycerol (10%), bromophenol blue (0.00025%), pH 8.6
Tris-acetate Native sample buffer: Tris HCl (100 mM), glycerol (10%), bromophenol blue (0.00025%), pH 8.6
IEF IEF sample buffer pH 3-7: Lysine (40 mM), glycerol (15%)
IEF sample buffer pH 3-10: Arginine (20 mM), Lysine (20 mM), glycerol (15%)
NativePAGE Bis-Tris NativePAGE Sample Buffer (4X): Bis-Tris (50 mM), 6 N HCl, NaCl (50 mM), Glycerol (10%), Ponceau S (0.001%), pH 7.2
Find native sample buffer recipes in gel-specific product manuals in the documents tab.

SDS running buffers

In protein electrophoresis (discontinuous buffer system), the primary anion in the gel is different (or discontinuous) from the primary anion in the running buffer. The tables below have the recommended SDS running buffers or native running buffers for each of the different gel chemistry buffering systems.

Gel system SDS running buffer
Tris-glycine Tris-glycine SDS: Tris base (25 mM), glycine (192 mM), SDS (0.1%), pH 8.3
Bis-Tris MES SDS: MES (50 mM), Tris base (50 mM), SDS (0.1%), EDTA (1 mM), pH 7.3
MOPS SDS: MOPS (50 mM), Tris base (50 mM), SDS (0.1%), EDTA (1 mM), pH 7.7
Tris-acetate Tris-acetate SDS: Tris base (50 mM), Tricine (50 mM), SDS (0.1%), pH 8.24
Tris-tricine Tricine-SDS: Tris base (100 mM), tricine (100 mM), SDS (0.1%), pH 8.3

Native running buffers

Gel system Native running buffer
Tris-glycine Tris-glycine native buffer: Tris base (25 mM), glycine (192 mM), pH 8.3
Tris-acetate Tris-glycine native buffer: Tris base (25 mM), glycine (192 mM), pH 8.3
IEF IEF cathode buffer pH 3-7: Lysine (40 mM)
IEF cathode buffer pH 3-10: Arginine (20 mM), lysine (20 mM)
IEF anode buffer: phosphoric acid 85% (7 mM)
Zymogram Tris-glycine SDS: Tris base (25 mM), glycine (192 mM), SDS (0.1%), pH 8.3
Find SDS running buffer recipes and native running buffer recipes in gel specific product manuals in the documents tab.

NuPAGE Bis-Tris SDS buffers & reagents

Protein sample Gel chemistry Sample buffers Running buffer Transfer buffer
Broad-range MW (6-400 kDa) Bis-tris Denaturing: LDS sample buffer

Reducing: Sample reducing agent
Small to medium-sized proteins (6–260 kDa): MES SDS buffer

Medium to large-size proteins (14–260 kDa): MOPs SDS buffer

Reduced samples: Antioxidant
Bis-tris transfer buffer

Bolt Bis-Tris Plus SDS buffers & reagents

Protein sample Gel chemistry Sample buffers Running buffer Transfer buffer
Broad-range MW (6-400 kDa) Bis-tris Denaturing: LDS sample buffer

Reducing: Sample reducing agent
Small to medium-sized proteins (6–260 kDa): MES SDS buffer

Medium to large-size proteins (14–260 kDa): MOPs SDS buffer

Reduced samples: Antioxidant
Bis-tris transfer buffer

Why LDS sample buffers

Bolt and NuPAGE LDS Sample Buffers are formulated with Coomassie G250 and phenol red as tracking dyes instead of bromophenol blue. Coomassie (SERVA) G250 gives a sharp dye front with both MES and MOPS SDS running buffers and migrates much closer to the moving ion front than bromophenol blue. Bromophenol blue runs more slowly than some peptides with the MES SDS Running Buffer. This ensures that small peptides do not run off the gel. The concentration of the tracking dye (Coomassie G250) is increased in Bolt and NuPAGE LDS Sample Buffers to enhance viewing of the dye front. LDS is used instead of SDS to allow the formulation of a 4X solution. The 4X solution provides the ability to have higher protein concentrations and less dilution of your protein sample.

Tris-glycine buffers & reagents

Protein sample Gel chemistry Sample buffers Running buffer Transfer buffer
Broad-range MW (6-400 kDa) Tris-glycine Denaturing: Tris-glycine SDS sample buffer

Native: Tris-glycine native sample buffer
Denaturing: Tris-glycine SDS buffer

Native: Tris-glycine native buffer
Tris-glycine transfer buffer

Tris-acetate buffers & reagents

Protein sample Gel chemistry Sample buffers Running buffer Transfer buffer
High MW
(40-500 kDa)
Tris-acetate Denaturing: LDS sample buffer

Native: Tris-glycine native sample buffer
Denaturing: Tris-acetate SDS buffer

Native: Tris-glycine native buffer
Bis-tris transfer buffer

Tricine SDS buffers & reagents

Protein sample Gel chemistry Sample buffers Running buffer Transfer buffer
Low MW
(2.5-40 kDa)
Tricine Denaturing: Tricine SDS sample buffer Tricine SDS buffer Tris-glycine transfer buffer

IEF gel recommended buffers

Zymography gel buffers & reagents

Gel type Sample buffer Running buffer Gel development buffer Gel renaturing buffer
Zymogram gel Tris-glycine SDS sample buffer Tris-glycine SDS running buffer Zymogram development buffer Zymogram renaturing buffer

NativePAGE Bis-Tris gel buffers & reagents

Gel casting

When pouring your own polyacrylamide gels, choosing high-quality reagents can help you achieve optimal electrophoresis results. SureCast buffers and reagents make it quick and easy to make high quality handcast Tris-glycine gels.

SDS-PAGE gel recipe for pour-your-own Tris-glycine gels

Prepare resolving gel solution according to the volumes in the following table. The volumes provided in the table are for a single gel. Scale volumes proportionally based on the number of gels to be cast.

  Polyacrylamide %
Solution 4% 6% 8% 10% 12% 14% 16% 18% 20%
SureCast Acrylamide (40%) 0.8mL 1.2mL 1.6mL 2.0mL 2.4mL 2.8mL 3.3mL 3.6mL 4.0mL
SureCast Resolving Buffer 2.0mL 2.0mL 2.0mL 2.0mL 2.0mL 2.0mL 2.0mL 2.0mL 2.0mL
Distilled water 5.1mL 4.7mL 4.3mL 3.9mL 3.5mL 3.1mL 2.7mL 2.3mL 1.9mL
10% SureCast Ammonium Persulfate (APS) 80µL 80µL 80µL 80µL 80µL 80µL 80µL 80µL 80µL
SureCast TEMED** 8µL 8µL 8µL 8µL 8µL 8µL 8µL 8µL 8µL

**Add this last and mix well just before the gel is to be poured

Prepare stacking gel solution according to the following table. The volumes provided in the table are for a single gel. Scale volumes proportionally based on the number of gels to be cast. Note: Solutions do not require degassing.

Solution 4%
SureCast Acrylamide (40%) 0.30 mL
SureCast Stacking Buffer 0.75 mL
Distilled water 1.92 mL
10% SureCast Ammonium Persulfate (APS) 30 µL
SureCast TEMED*** 3 µL

***Add this last and mix well just before the gel is to be poured

Learn more about gel casting and find additional SDS-PAGE gel recipes.

Ordering quick links

NuPAGE Bis-Tris Buffers

Bolt Bis-Tris Plus Buffers

Tris-Glycine Buffers

Tris-Acetate Buffers

Tricine System Buffers

IEF Buffers

Zymogram Buffers

NativePAGE Bis-Tris Buffers

Handcast Buffers & Reagents

SDS sample buffers or loading buffers

Protein samples prepared for SDS-PAGE analysis are denatured by heating in the presence of a sample buffer containing 1–2% SDS or LDS with or without a reducing agent such as 20 mM DTT, 2-mercaptoethanol (BME) or TCEP. Sample buffers or loading buffers also contain glycerol so that they are heavier than water and sink neatly to the bottom of the buffer-submerged well when added to a gel. In addition, a negatively charged, low-molecular weight dye is also included in the sample buffer that will migrate at the buffer-front, enabling one to monitor the progress of electrophoresis. The most common tracking dyes for sample loading buffers are bromophenol blue, phenol red, and Coomassie blue. The table below summaries common sample buffers used in the different gel buffer systems.

Gel system Denaturing sample buffer (final concentrations) Sample preparation
Tris-glycine Tris-glycine SDS sample buffer: Tris HCl (63 mM), glycerol (10%), SDS (2%), bromophenol blue (0.0025%), pH 6.8 Heat samples at 85°C for 2–5 minutes with sample buffer
Bis-Tris LDS sample buffer: Tris base (141 mM), Tris HCl (106 mM), LDS (2%), EDTA (0.51 mM), SERVA Blue G-250 (0.22 mM), phenol red (0.175 mM), pH 8.5 Heat samples at 70°C for 10 minutes with sample buffer
Tris-Acetate LDS sample buffer: Tris base (141 mM), Tris HCl (106 mM), LDS (2%), EDTA (0.51 mM), SERVA Blue G-250 (0.22 mM), phenol red (0.175 mM), pH 8.5 Heat samples at 70°C for 10 minutes with sample buffer
Tris-Tricine Tricine SDS sample buffer: Tris HCl (450 mM), glycerol (12%), SDS (4%), Coomassie Blue G (0.00075%), phenol red (0.0025%), pH 8.45 Heat samples at 85°C for 2–5 minutes with sample buffer
Zymogram Tris-glycine SDS: Tris HCl (63 mM), glycerol (10%), SDS (2%), bromophenol blue (0.0025%), pH 6.8 Do not heat samples, load immediately onto gel after adding to sample

For fluorescence applications, we recommend that you use sample buffers that do not contain dyes that interfere in fluorescent channels (e.g., bromophenol blue). Fluorescent Compatible Sample Buffer does not contain bromophenol blue or other interfering dyes and can be used with reducing or nonreducing SDS-PAGE applications.

Find SDS sample buffer recipes in gel-specific product manuals in the documents tab.

Native sample buffers

Protein samples prepared for native PAGE applications are prepared in sample buffers or loading buffers that do not contain detergents to maintain the native state of the proteins. The loading buffers contain glycerol so that they are heavier than water and sink neatly to the bottom of the buffer-submerged well when added to a gel. In addition, a negatively charged, low-molecular weight dye is also included in the sample buffer that will migrate at the buffer-front, enabling one to monitor the progress of electrophoresis.

When preparing samples for native applications, protein samples should not be heated and loaded immediately onto the gel after mixing with the sample buffer.

Gel system Native sample buffer (final concentrations)
Tris-glycine Native sample buffer: Tris HCl (100 mM), glycerol (10%), bromophenol blue (0.00025%), pH 8.6
Tris-acetate Native sample buffer: Tris HCl (100 mM), glycerol (10%), bromophenol blue (0.00025%), pH 8.6
IEF IEF sample buffer pH 3-7: Lysine (40 mM), glycerol (15%)
IEF sample buffer pH 3-10: Arginine (20 mM), Lysine (20 mM), glycerol (15%)
NativePAGE Bis-Tris NativePAGE Sample Buffer (4X): Bis-Tris (50 mM), 6 N HCl, NaCl (50 mM), Glycerol (10%), Ponceau S (0.001%), pH 7.2
Find native sample buffer recipes in gel-specific product manuals in the documents tab.

SDS running buffers

In protein electrophoresis (discontinuous buffer system), the primary anion in the gel is different (or discontinuous) from the primary anion in the running buffer. The tables below have the recommended SDS running buffers or native running buffers for each of the different gel chemistry buffering systems.

Gel system SDS running buffer
Tris-glycine Tris-glycine SDS: Tris base (25 mM), glycine (192 mM), SDS (0.1%), pH 8.3
Bis-Tris MES SDS: MES (50 mM), Tris base (50 mM), SDS (0.1%), EDTA (1 mM), pH 7.3
MOPS SDS: MOPS (50 mM), Tris base (50 mM), SDS (0.1%), EDTA (1 mM), pH 7.7
Tris-acetate Tris-acetate SDS: Tris base (50 mM), Tricine (50 mM), SDS (0.1%), pH 8.24
Tris-tricine Tricine-SDS: Tris base (100 mM), tricine (100 mM), SDS (0.1%), pH 8.3

Native running buffers

Gel system Native running buffer
Tris-glycine Tris-glycine native buffer: Tris base (25 mM), glycine (192 mM), pH 8.3
Tris-acetate Tris-glycine native buffer: Tris base (25 mM), glycine (192 mM), pH 8.3
IEF IEF cathode buffer pH 3-7: Lysine (40 mM)
IEF cathode buffer pH 3-10: Arginine (20 mM), lysine (20 mM)
IEF anode buffer: phosphoric acid 85% (7 mM)
Zymogram Tris-glycine SDS: Tris base (25 mM), glycine (192 mM), SDS (0.1%), pH 8.3
Find SDS running buffer recipes and native running buffer recipes in gel specific product manuals in the documents tab.

NuPAGE Bis-Tris SDS buffers & reagents

Protein sample Gel chemistry Sample buffers Running buffer Transfer buffer
Broad-range MW (6-400 kDa) Bis-tris Denaturing: LDS sample buffer

Reducing: Sample reducing agent
Small to medium-sized proteins (6–260 kDa): MES SDS buffer

Medium to large-size proteins (14–260 kDa): MOPs SDS buffer

Reduced samples: Antioxidant
Bis-tris transfer buffer

Bolt Bis-Tris Plus SDS buffers & reagents

Protein sample Gel chemistry Sample buffers Running buffer Transfer buffer
Broad-range MW (6-400 kDa) Bis-tris Denaturing: LDS sample buffer

Reducing: Sample reducing agent
Small to medium-sized proteins (6–260 kDa): MES SDS buffer

Medium to large-size proteins (14–260 kDa): MOPs SDS buffer

Reduced samples: Antioxidant
Bis-tris transfer buffer

Why LDS sample buffers

Bolt and NuPAGE LDS Sample Buffers are formulated with Coomassie G250 and phenol red as tracking dyes instead of bromophenol blue. Coomassie (SERVA) G250 gives a sharp dye front with both MES and MOPS SDS running buffers and migrates much closer to the moving ion front than bromophenol blue. Bromophenol blue runs more slowly than some peptides with the MES SDS Running Buffer. This ensures that small peptides do not run off the gel. The concentration of the tracking dye (Coomassie G250) is increased in Bolt and NuPAGE LDS Sample Buffers to enhance viewing of the dye front. LDS is used instead of SDS to allow the formulation of a 4X solution. The 4X solution provides the ability to have higher protein concentrations and less dilution of your protein sample.

Tris-glycine buffers & reagents

Protein sample Gel chemistry Sample buffers Running buffer Transfer buffer
Broad-range MW (6-400 kDa) Tris-glycine Denaturing: Tris-glycine SDS sample buffer

Native: Tris-glycine native sample buffer
Denaturing: Tris-glycine SDS buffer

Native: Tris-glycine native buffer
Tris-glycine transfer buffer

Tris-acetate buffers & reagents

Protein sample Gel chemistry Sample buffers Running buffer Transfer buffer
High MW
(40-500 kDa)
Tris-acetate Denaturing: LDS sample buffer

Native: Tris-glycine native sample buffer
Denaturing: Tris-acetate SDS buffer

Native: Tris-glycine native buffer
Bis-tris transfer buffer

Tricine SDS buffers & reagents

Protein sample Gel chemistry Sample buffers Running buffer Transfer buffer
Low MW
(2.5-40 kDa)
Tricine Denaturing: Tricine SDS sample buffer Tricine SDS buffer Tris-glycine transfer buffer

IEF gel recommended buffers

Zymography gel buffers & reagents

Gel type Sample buffer Running buffer Gel development buffer Gel renaturing buffer
Zymogram gel Tris-glycine SDS sample buffer Tris-glycine SDS running buffer Zymogram development buffer Zymogram renaturing buffer

NativePAGE Bis-Tris gel buffers & reagents

Gel casting

When pouring your own polyacrylamide gels, choosing high-quality reagents can help you achieve optimal electrophoresis results. SureCast buffers and reagents make it quick and easy to make high quality handcast Tris-glycine gels.

SDS-PAGE gel recipe for pour-your-own Tris-glycine gels

Prepare resolving gel solution according to the volumes in the following table. The volumes provided in the table are for a single gel. Scale volumes proportionally based on the number of gels to be cast.

  Polyacrylamide %
Solution 4% 6% 8% 10% 12% 14% 16% 18% 20%
SureCast Acrylamide (40%) 0.8mL 1.2mL 1.6mL 2.0mL 2.4mL 2.8mL 3.3mL 3.6mL 4.0mL
SureCast Resolving Buffer 2.0mL 2.0mL 2.0mL 2.0mL 2.0mL 2.0mL 2.0mL 2.0mL 2.0mL
Distilled water 5.1mL 4.7mL 4.3mL 3.9mL 3.5mL 3.1mL 2.7mL 2.3mL 1.9mL
10% SureCast Ammonium Persulfate (APS) 80µL 80µL 80µL 80µL 80µL 80µL 80µL 80µL 80µL
SureCast TEMED** 8µL 8µL 8µL 8µL 8µL 8µL 8µL 8µL 8µL

**Add this last and mix well just before the gel is to be poured

Prepare stacking gel solution according to the following table. The volumes provided in the table are for a single gel. Scale volumes proportionally based on the number of gels to be cast. Note: Solutions do not require degassing.

Solution 4%
SureCast Acrylamide (40%) 0.30 mL
SureCast Stacking Buffer 0.75 mL
Distilled water 1.92 mL
10% SureCast Ammonium Persulfate (APS) 30 µL
SureCast TEMED*** 3 µL

***Add this last and mix well just before the gel is to be poured

Learn more about gel casting and find additional SDS-PAGE gel recipes.

Ordering quick links

NuPAGE Bis-Tris Buffers

Bolt Bis-Tris Plus Buffers

Tris-Glycine Buffers

Tris-Acetate Buffers

Tricine System Buffers

IEF Buffers

Zymogram Buffers

NativePAGE Bis-Tris Buffers

Handcast Buffers & Reagents

Resources