Invitrogen NuPAGE Tris-Acetate Gels are high-performance gels that have been optimized for separating proteins of large molecular weight.
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 | 2 - 8°C |
| 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 | SureLock Tandem Midi Gel Tank, 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.
Tris-acetate gels enable the best separation of HMW proteins. (A) Optimal results are obtained in the yellow shaded areas. (B) Better transfer is seen using the tris-acetate gel over a 4–20% Tris-glycine gel: 9 ng is visualized on the Tris-acetate gel versus 620 ng visualized on the tris-glycine gradient gel.
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 |
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 | 2 - 8°C |
| 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 | SureLock Tandem Midi Gel Tank, 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.
Tris-acetate gels enable the best separation of HMW proteins. (A) Optimal results are obtained in the yellow shaded areas. (B) Better transfer is seen using the tris-acetate gel over a 4–20% Tris-glycine gel: 9 ng is visualized on the Tris-acetate gel versus 620 ng visualized on the tris-glycine gradient gel.
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 |
For Research Use Only. Not for use in diagnostic procedures.
