NuPAGE Tris-Acetate Gels

Invitrogen NuPAGE Tris-Acetate Gels are high-performance gels that have been optimized for separating proteins of large molecular weight.

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NuPAGE Tris-Acetate Gel specifications

Available gel sizes Mini: 8 cm x 8 cm (1 or 1.5 mm thick)
Midi: 8 cm x 13 cm (1 mm thick)
Storage conditions Room temperature
Shelf life 8 months
Recommended sample buffer SDS-PAGE: NuPAGE LDS Sample Buffer
Native-PAGE: Novex Tris-Glycine Native Sample Buffer
Recommended running buffer SDS-PAGE: NuPAGE Tris-Acetate SDS Running Buffer
Native-PAGE: Novex Tris-Glycine Native Running Buffer
Recommended transfer buffer NuPAGE Transfer Buffer
Gel chemistry Tris-acetate
Available polyacrylamide concentrations 7%, 3–8%
Separation range (denaturing) 30–500 kDa
For use with (equipment) mini gels Mini Gel Tank or XCell SureLock Mini-Cell
For use with (equipment) midi gels Invitrogen XCell4 SureLock Midi-Cell or Bio-Rad Criterion (with adapters only)
Mode of separation Molecular weight
Applications SDS-PAGE, Native-PAGE
Well type* Mini: 10, 12, 15-wells
Midi: 12+2, 20, 26-wells

*Not all percentages are available in every well type

Reliable separation and transfer of high molecular weight (HMW) proteins is a common challenge for life science researchers. Choosing the right gel is a key factor in the successful transfer of HMW proteins. A popular general-use gel is a 4–20% Tris-glycine gel, which can effectively separate a mixed range of proteins. However, HMW proteins will be compressed into a narrow region at the top of the gel. A better option for HMW proteins is a Tris-acetate gel or a low percentage non-gradient Tris-glycine or Bis-Tris gel. When specially targeting HMW proteins, optimal transfer can be achieved with a Tris-acetate gel.

Tris-acetate gels maintain a neutral pH and separate HMW proteins with higher resolution than Bis-Tris or Tris-glycine gels. Comparison of HMW protein separation using different gel chemistries and gradients shows best separation and resolution of HMW proteins using a 3–8% Tris-acetate gel. This increased resolution leads to increased transfer efficiencies and higher sensitivity.

NuPAGE Tris-Acetate Gels are designed to give optimal separation of large molecular weight proteins during gel electrophoresis. In comparison to traditional Tris-glycine SDS-PAGE gels, NuPAGE Tris-Acetate Gels have an environment of pH 8.1, which minimizes protein modifications and results in sharper bands.

 Features of NuPAGE Tris-Acetate Gels include:

  • High resolution—gels offer optimal separation of high molecular weight proteins
  • Better protein integrity—sample preparation process has been optimized to help preserve your proteins
  • Longer shelf life—gels can be stored for at least 8 months

NuPAGE Tris-Acetate Gels do not contain SDS and so can be used to separate proteins in native or denatured form. For denatured proteins, we recommend using LDS sample buffer and tris-acetate SDS running buffer. For native proteins, traditional Tris-glycine native sample buffer and a Tris-glycine native running buffer should be used.

The NuPAGE Tris-Acetate Gels use a discontinuous buffer system involving three ions:

  • Acetate (–) is from the gel buffer and serves as a leading ion due to its high affinity to the anode relative to other anions in the system. The gel buffer ions are tris (+) and acetate (–) (pH 7.0).
  • Tricine (–) is from the running buffer and serves as the trailing ion. The running buffer ions are tris (+), tricine (–) and dodecylsulfate (pH 8.3).
  • Tris (+) is the common ion present in the gel buffer and running buffer. The Tris-acetate system also operates at a significantly lower operating pH of 8.1 during electrophoresis. 

Better sample integrity

Sample preparation is a crucial step in successful protein analysis. The pH of the traditional Laemmli-style sample buffer changes from 6.8 to 5.2 when heated at 100°C. This lower pH is known to induce aspartyl-prolyl (Asp-Pro) peptide bond cleavage, which leads to protein degradation. By maintaining a >7.0 pH environment, the Invitrogen NuPAGE LDS Sample Buffer preserves protein integrity by minimizing this Asp-Pro cleavage. Sample integrity is maintained throughout electrophoresis with the Invitrogen NuPAGE system; protein degradation is seen in samples prepared with Laemmli (tris-glycine) sample buffer. The Invitrogen NuPAGE Antioxidant running buffer additive greatly minimizes protein oxidation during electrophoresis and keeps reduced protein bands sharp and clear.

Currently using Tris-glycine or Tricine gels? Use the NuPAGE Gel conversion guide below to find the appropriate NuPAGE gel and corresponding optimized buffer.

NuPAGE Gel conversion guide

If you are using: Then we recommend NuPAGE gel: Buffer system
Tris-Glycine gel
4% NuPAGE 3-8% Tris-Acetate Tris-Acetate
6% NuPAGE 3-8% Tris-Acetate Tris-Acetate
10% NuPAGE 8% Bis-Tris MOPS
NuPAGE 10% Bis-Tris MOPS
12% NuPAGE 10% Bis-Tris MOPS
14% NuPAGE 12% Bis-Tris MOPS
16% NuPAGE 12% Bis-Tris MES
18% NuPAGE 12% Bis-Tris MES
4-12% NuPAGE 3-8% Tris-Acetate Tris-Acetate
NuPAGE 4-12% Bis-Tris MOPS
4-20% NuPAGE 4-12% Bis-Tris MES
8-16% NuPAGE 4-12% Bis-Tris MOPS
10-20% NuPAGE 12% Bis-Tris MOPS
Tricine gel
10% NuPAGE 8% Bis-Tris MES
NuPAGE 10% Bis-Tris MES
16% NuPAGE 4-12% Bis-Tris MES
10-20% NuPAGE 4-12% Bis-Tris MES
Migration charts for NuPAGE Tris-Acetate Gels

NuPAGE Tris-Acetate Gel specifications

Available gel sizes Mini: 8 cm x 8 cm (1 or 1.5 mm thick)
Midi: 8 cm x 13 cm (1 mm thick)
Storage conditions Room temperature
Shelf life 8 months
Recommended sample buffer SDS-PAGE: NuPAGE LDS Sample Buffer
Native-PAGE: Novex Tris-Glycine Native Sample Buffer
Recommended running buffer SDS-PAGE: NuPAGE Tris-Acetate SDS Running Buffer
Native-PAGE: Novex Tris-Glycine Native Running Buffer
Recommended transfer buffer NuPAGE Transfer Buffer
Gel chemistry Tris-acetate
Available polyacrylamide concentrations 7%, 3–8%
Separation range (denaturing) 30–500 kDa
For use with (equipment) mini gels Mini Gel Tank or XCell SureLock Mini-Cell
For use with (equipment) midi gels Invitrogen XCell4 SureLock Midi-Cell or Bio-Rad Criterion (with adapters only)
Mode of separation Molecular weight
Applications SDS-PAGE, Native-PAGE
Well type* Mini: 10, 12, 15-wells
Midi: 12+2, 20, 26-wells

*Not all percentages are available in every well type

Reliable separation and transfer of high molecular weight (HMW) proteins is a common challenge for life science researchers. Choosing the right gel is a key factor in the successful transfer of HMW proteins. A popular general-use gel is a 4–20% Tris-glycine gel, which can effectively separate a mixed range of proteins. However, HMW proteins will be compressed into a narrow region at the top of the gel. A better option for HMW proteins is a Tris-acetate gel or a low percentage non-gradient Tris-glycine or Bis-Tris gel. When specially targeting HMW proteins, optimal transfer can be achieved with a Tris-acetate gel.

Tris-acetate gels maintain a neutral pH and separate HMW proteins with higher resolution than Bis-Tris or Tris-glycine gels. Comparison of HMW protein separation using different gel chemistries and gradients shows best separation and resolution of HMW proteins using a 3–8% Tris-acetate gel. This increased resolution leads to increased transfer efficiencies and higher sensitivity.

NuPAGE Tris-Acetate Gels are designed to give optimal separation of large molecular weight proteins during gel electrophoresis. In comparison to traditional Tris-glycine SDS-PAGE gels, NuPAGE Tris-Acetate Gels have an environment of pH 8.1, which minimizes protein modifications and results in sharper bands.

 Features of NuPAGE Tris-Acetate Gels include:

  • High resolution—gels offer optimal separation of high molecular weight proteins
  • Better protein integrity—sample preparation process has been optimized to help preserve your proteins
  • Longer shelf life—gels can be stored for at least 8 months

NuPAGE Tris-Acetate Gels do not contain SDS and so can be used to separate proteins in native or denatured form. For denatured proteins, we recommend using LDS sample buffer and tris-acetate SDS running buffer. For native proteins, traditional Tris-glycine native sample buffer and a Tris-glycine native running buffer should be used.

The NuPAGE Tris-Acetate Gels use a discontinuous buffer system involving three ions:

  • Acetate (–) is from the gel buffer and serves as a leading ion due to its high affinity to the anode relative to other anions in the system. The gel buffer ions are tris (+) and acetate (–) (pH 7.0).
  • Tricine (–) is from the running buffer and serves as the trailing ion. The running buffer ions are tris (+), tricine (–) and dodecylsulfate (pH 8.3).
  • Tris (+) is the common ion present in the gel buffer and running buffer. The Tris-acetate system also operates at a significantly lower operating pH of 8.1 during electrophoresis. 

Better sample integrity

Sample preparation is a crucial step in successful protein analysis. The pH of the traditional Laemmli-style sample buffer changes from 6.8 to 5.2 when heated at 100°C. This lower pH is known to induce aspartyl-prolyl (Asp-Pro) peptide bond cleavage, which leads to protein degradation. By maintaining a >7.0 pH environment, the Invitrogen NuPAGE LDS Sample Buffer preserves protein integrity by minimizing this Asp-Pro cleavage. Sample integrity is maintained throughout electrophoresis with the Invitrogen NuPAGE system; protein degradation is seen in samples prepared with Laemmli (tris-glycine) sample buffer. The Invitrogen NuPAGE Antioxidant running buffer additive greatly minimizes protein oxidation during electrophoresis and keeps reduced protein bands sharp and clear.

Currently using Tris-glycine or Tricine gels? Use the NuPAGE Gel conversion guide below to find the appropriate NuPAGE gel and corresponding optimized buffer.

NuPAGE Gel conversion guide

If you are using: Then we recommend NuPAGE gel: Buffer system
Tris-Glycine gel
4% NuPAGE 3-8% Tris-Acetate Tris-Acetate
6% NuPAGE 3-8% Tris-Acetate Tris-Acetate
10% NuPAGE 8% Bis-Tris MOPS
NuPAGE 10% Bis-Tris MOPS
12% NuPAGE 10% Bis-Tris MOPS
14% NuPAGE 12% Bis-Tris MOPS
16% NuPAGE 12% Bis-Tris MES
18% NuPAGE 12% Bis-Tris MES
4-12% NuPAGE 3-8% Tris-Acetate Tris-Acetate
NuPAGE 4-12% Bis-Tris MOPS
4-20% NuPAGE 4-12% Bis-Tris MES
8-16% NuPAGE 4-12% Bis-Tris MOPS
10-20% NuPAGE 12% Bis-Tris MOPS
Tricine gel
10% NuPAGE 8% Bis-Tris MES
NuPAGE 10% Bis-Tris MES
16% NuPAGE 4-12% Bis-Tris MES
10-20% NuPAGE 4-12% Bis-Tris MES
Migration charts for NuPAGE Tris-Acetate Gels
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