UltraBio™ Tris-Acetate Precast PAGE Gel for High Molecular Weight Protein

Item Number

T753564

• Product Name: UltraBio™ Tris-Acetate Precast PAGE Gel for High Molecular Weight Protein

Product Description

The UltraBio™ Tris-Acetate Precast PAGE Gel for High Molecular Weight Protein is a safe, high-quality, and ready-to-use polyacrylamide gel primarily used to analyze high molecular weight proteins (40-500kDa). It can be used for both denaturing and native protein electrophoresis with the Tris-Acetate SDS (SDS-PAGE) and Tris-Glycine (Native-PAGE) running buffers, respectively. It takes approximately 60 minutes to complete electrophoresis, and well-separated and sharp protein bands can be obtained. This precast gel has a 1.5cm high 4% stacking gel, with a very good protein separation effect. Moreover, this product uses glass plates to reduce non-specific adsorptions. This product has identical or even better performance than home-made PAGE gels and can be routinely used for PAGE and Western analysis.

Aladdin provides both UltraBio™ precast gradient/fixed gels of different concentrations with 10 or 15 loading wells. The gradient gel concentration is 3-8%, and the fixed gel concentration is 7%. Please refer to the table below for the optimal protein separation range for the UltraBio™ Tris-Acetate Precast PAGE Gels.

Product No.	Gel Concentration	Number of Wells	Maximum Loading Volume	Electrophoresis Buffer System	Membrane Transfer Buffer System	Optimal Separation Range
T753564-A1-10EA	7%	10	60μl	Tris-Acetate SDS or Tris-Glycine	Tris-Glycine	40-500kD
T753564-B1-10EA	7%	15	30μl	Tris-Acetate SDS or Tris-Glycine	Tris-Glycine	40-500kD
T753564-A2-10EA	3-8%	10	60μl	Tris-Acetate SDS or Tris-Glycine	Tris-Glycine	40-500kD
T753564-B2-10EA	3-8%	15	30μl	Tris-Acetate SDS or Tris-Glycine	Tris-Glycine	40-500kD

This product has a 4% stacking gel with a height of 1.5cm to ensure very sharp protein bands can be obtained.

The ratio of acrylamide to bisacrylamide is 29:1, and the gel thickness is 1.5mm. The maximum loading volume per well of 10-well and 15-well gels is 60μl and 30μl, respectively. The plate size is 98×84×4.1mm (width × height × thickness), with a gel size of 81×74×1.5mm (width × height × thickness).

Polyacrylamide gel electrophoresis (PAGE) is a widely used technique in life science research for the separation, purification, assay, identification, and molecular weight analysis of proteins, nucleic acids, and protein-nucleic acid complexes. The commonly used Western blot technique is based on PAGE. Glycine-SDS-PAGE (also known as Laemmli-SDS-PAGE, based on the Tris-Glycine buffer system) and Tricine-SDS-PAGE (based on the Tris-Tricine buffer system) are widely used methods for separating proteins based on their molecular weight. Tricine-SDS-PAGE is particularly effective in separating proteins smaller than 30kDa, while Glycine-SDS-PAGE is recommended for proteins larger than 30kDa. The separation characteristics of proteins in different systems are related to the pKa values of Glycine and Tricine. Proteins of different sizes have different electrophoretic mobilities in different buffer systems, and proteins within specific size ranges can be resolved with high resolution. Therefore, when separating high molecular weight proteins, it is important to choose the appropriate electrophoresis system. It is also recommended to use gradient gels with decreasing acrylamide concentration from the bottom to the top. Currently, the Tris-Acetate gel system is commonly used for separating high molecular weight proteins. In the Tris-Acetate gel system, Acetate (-) comes from the gel buffer, and compared to other anions in the system, Acetate (-) has a high affinity for the anode and can serve as the leading ion. The gel buffer contains Tris (+) and Acetate (-) at a pH of 7.0, which provides high resolution in the gel. The electrophoresis buffer contains Tris (+), Tricine (-), and dodecyl sulfate (DS), with Tricine (-) acting as the trailing ion. The pH of the electrophoresis buffer is 8.24, which reduces protein modifications and results in sharper bands. The results of separation of high molecular weight proteins have also shown that Tris-Acetate gels provide good separation, high transfer efficiency, high sensitivity and excellent resolution. In addition, Tris-Acetate gels have been reported to serve as a stable and cost-effective tool for the study of protein oligomerization, providing excellent protein oligomerization results using only a single gel.

Unlike traditional Tris-Glycine gels, UltraBio™ Tris-Acetate Precast PAGE Gels are prepared with neutral pH tris-acetate buffer and do not contain SDS. They can be used for both denaturing and native protein electrophoresis.

After electrophoresis, protein can be transferred from the gel to blot membrane with the Tris-Glycine buffer system.However, the concentration of ethanol or methanol in the transfer buffer needs to be reduced to 5%.

Selection of 10-well and 15-well precast gels: Protein bands obtained by 10-well precast gels are flatter and sharper. When there is a large number of samples to be examined or protein quantification is required, we recommend using the 15-well precast gel which offers greater throughput and is more convenient for statistical quantitation analysis.

This product is safe and convenient to use. There is no need for gel preparation and casting, thus avoiding contact with toxic and irritating reagents. After electrophoresis, the gel can be easily removed from the glass plates.

The quality of this product is stable, with high consistency between batches.

This product has good protein resolution performance. Proteins in the gel can be transferred efficiently as well.

The electrophoresis time is short. The denaturing electrophoresis can be completed in 50-70 minutes at 150V, while the native electrophoresis can be completed in 90-180 minutes at 150V. The run time varies depending on experimental requirements.

Precautions:

Do NOT freeze this product at temperatures below 0℃. 

The electrophoresis buffer in the inner chamber of the electrophoresis tank and the transfer buffer should be freshly prepared. Low-quality buffers, recycled buffers, or buffers that have been kept for too long period of time will result in poor protein resolution and transfer. 

After electrophoresis, protein transfer can be performed using the Tris-Glycine transfer buffer system. Ethanol or methanol can cause precipitation of large protein molecules, which can be avoided by reducing the percentage of ethanol or methanol in the transfer buffer to 5%. SDS can be additionally added to a final concentration of 0.1% to further ensure no precipitation of proteins. SDS enables a uniform negative charge of proteins, allowing them to be more easily transferred from the gel to the membrane. 

For transfer, we recommend using 0.45μm PVDF Membrane.

This product is for R&D only. Not for drug, household, or other uses.

For your safety and health, please wear a lab coat and disposable gloves during the operation.