Tris CAPS buffer 10X
Cat No.: DCP-TCAPS10X
Size: 500 ml & 1 L
Sterile; Ultrapure, Filtered 0.1-micron membrane TWICE and 0.04-micron membrane ONCE.
Description
Tris-CAPS Transfer Buffer is a specialized buffer system used primarily in semi-dry blotting for Western blotting applications. This buffer is particularly advantageous for transferring proteins efficiently from polyacrylamide gels to membranes. In semi-dry blotting, the Tris-CAPS buffer is used in a discontinuous system where different buffers are applied to the anode and cathode sides. Methanol is used on the anode side to promote protein binding to the membrane, while SDS is used on the cathode side to facilitate protein migration out of the gel. Methanol helps strip SDS from proteins, enhancing their binding to membranes, but can also shrink gels and membranes, affecting the transfer of large proteins. SDS aids in protein migration but can inhibit binding if used excessively. Like all transfer buffers, the Tris-CAPS system may require optimization depending on the specific proteins and membranes used, as well as the specific setup of the semi-dry blotting apparatus.
Methanol: Typically 15% (v/v) needs to be added to the final concentration on the anode side.
SDS: 0.1% needs to be added to the final concentration on the cathode side to promote protein migration out of the gel.
Applications
-
Semi-Dry Blotting: The Tris-CAPS buffer is specifically designed for semi-dry transfer methods. In this setup, the buffer is used in a discontinuous system where different buffers are applied to the anode and cathode sides. This allows for enhanced protein transfer efficiency by optimizing the conditions on each side of the blotting assembly.
-
Protein Transfer: The Tris-CAPS buffer system is used to facilitate the transfer of proteins from polyacrylamide gels to membranes. It is particularly effective for proteins with a broad range of molecular weights, ensuring efficient migration and binding.
-
Use with Basic Proteins: The higher pH of the Tris-CAPS buffer (approximately 9.6) makes it suitable for transferring basic proteins, which might not transfer as effectively with other buffer systems.
Advantages:
-
Enhanced Transfer Efficiency: The discontinuous buffer system allows for the use of different buffers on the anode and cathode sides of the blotting assembly. This setup optimizes the conditions for protein transfer by taking advantage of the distinct properties of each buffer component.
-
Optimization of Protein Elution and Binding: Methanol increases the protein's affinity for the membrane by removing SDS, which exposes more hydrophobic sites on the protein for binding. Conversely, SDS aids in the elution of proteins from the gel matrix, ensuring efficient transfer. By using these components in a discontinuous manner, the system capitalizes on the beneficial effects of each, improving overall transfer efficiency.
-
Flexibility and Customization: The ability to use different buffers on each side of the transfer stack allows for customization based on the specific proteins being transferred. This flexibility can lead to better optimization for proteins with varying sizes and properties, enhancing the quality of the transfer.
-
Reduced Buffer Volume Requirement: Semi-dry blotting systems, in general, require less buffer compared to wet transfer systems. The discontinuous Tris-CAPS system further optimizes this by using filter papers as buffer reservoirs, making the process more resource-efficient.
-
Rapid Transfer: Semi-dry blotting with a discontinuous buffer system typically allows for faster transfer times compared to wet transfer methods, making it suitable for high-throughput applications
Composition:
| Ingredients |
Concentration |
| Tris base |
60 mM |
| CAPS |
40 mM |
Quality Control:
-
Appearance: Clear, Colorless Liquid.
-
pH: 9.6
-
Sterility: Filtered 0.1 micron Twice and 0.04 micron once in a sterile environment.
-
DNase activity: None was detected after incubation of plasmid DNA and this product for 18 hr at room temperature.
-
RNase activity: No RNase activity was detected after incubation of ribosomal RNA and this product for 18 hr at room temperature.
Storage and Shelf Life:
-
Storage: 4 ℃
-
Shelf life: 1 year
CUSTOMIZATION: Please INQUIRE if other concentrations, additions of chemicals/compounds/proteins/supplements, different pH, and modifications are needed.
All DiagnoCine Precision Sterile buffers are filtered-sterilized with a 0.1-micron filtration two times and 0.04-micron filtration one time. Thus, mycoplasma contamination is prevented. The smallest size mycoplasma type can be about 0.2 microns.
* This product is manufactured under ISO 13485-certified and CE-approved facilities (Suppliers of DiagnoCine Precision). All final packaging, quality assurance, and testing are done at the DiagnoCine R&D and Quality Testing Center. All specific customization requests and assembly were accomplished at DiagnoCine Precision in Totowa, New Jersey, USA.
