Tris-HCl Buffer, Sterile [0.5X]
Cat No.: DCP-THB1.5X
Size: 1000 mL
Sterile; Ultrapure, Filtered 0.1-micron membrane TWICE and 0.04-micron membrane ONCE.
Description:
Tris-HCl buffer is widely used in molecular biology techniques like gel electrophoresis, nucleic acid work, protein extraction and purification.It can be used as stacking gel buffer in polyacrylamide gel electrophoresis (PAGE) for protein separation. It creates a pH discontinuity between the stacking and resolving gels, which helps concentrate proteins into sharp bands before entering the resolving gel. The lower pH (6.8) in the stacking gel allows proteins to stack tightly at the interface with the resolving gel.
Advantages:
- Improved resolution: The stacking gel buffer creates a discontinuous buffer system that significantly enhances protein resolution compared to continuous systems.
- Sample concentration: It allows for the loading of large volumes of dilute protein samples, concentrating them into narrow zones or stacks.
- Preservation of native protein structure: Without SDS, proteins maintain their native conformation and interactions, allowing for the study of protein complexes and oligomeric states.
- Retention of enzymatic activity: The absence of SDS helps preserve the functional properties of proteins, including enzymatic activity.
- Reduced protein modifications: The lower pH of SDS-free buffers (typically around 6.8) minimizes protein modifications during electrophoresis, resulting in sharper bands.
- Compatibility with downstream applications: SDS-free conditions are often more suitable for subsequent analyses that require native protein structures, such as activity assays or protein-protein interaction studies.
Application:
- SDS-PAGE: Stacking gel buffer is a crucial component in sodium dodecyl sulfate polyacrylamide gel electrophoresis, the most popular form of protein electrophoresis.
- Native PAGE: It can be used in native polyacrylamide gel electrophoresis to separate proteins while preserving their native conformation and activity.
- 2D electrophoresis: Stacking gel buffer is used in the second dimension of two-dimensional electrophoresis techniques.
- Protein molecular weight determination: The stacking effect contributes to more accurate sizing of proteins in SDS-PAGE.
- Analysis of protein complexes: In blue native PAGE, a variation of native PAGE, stacking gel buffer helps separate intact protein complexes.
- Preparative electrophoresis: The concentration effect of the stacking gel is useful when purifying proteins by electrophoresis.
- High-resolution protein separation: The stacking mechanism allows for the separation of thousands of polypeptides in a single slab gel when combined with other techniques like isoelectric focusing
Composition:
- 0.5M Tris base
- HCl to adjust the pH
Quality Control:
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Appearance: Clear, Colorless Liquid.
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pH: 6.8
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Sterility: Filtered 0.1 micron Twice + 0.04 micron in a sterile environment.
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DNase activity: None was detected after incubation of plasmid DNA and this product for 18 hr at room temperature.
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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:
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Storage: 4 ℃
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Shelf life: 2 years
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
