Bis-Tris Transfer Buffer
Cat No.: DCP-BTTB1X
Size: 500 mL & 1 L
Sterile; Ultrapure, Filtered 0.1-micron membrane TWICE and 0.04-micron membrane ONCE.
Description:
READY-to-USE 1X Bis-Tris Transfer Buffer is a buffer solution used for transferring proteins from polyacrylamide gels to membranes (e.g., nitrocellulose or PVDF) during Western blotting procedures. It is specifically formulated for use with Bis-Tris polyacrylamide gels, which are commonly used for separating proteins under denaturing conditions. The presence of EDTA chelates divalent metal ions, preventing their interference with the transfer process. Bis-Tris is non-toxic, unlike cacodylate which is highly toxic and carcinogenic. This makes Bis-Tris a safer choice for biological and biochemical applications. Bis-Tris has a lower UV absorption compared to maleic acid. Maleic acid absorbs strongly in the UV range, which can interfere with spectrophotometric assays and measurements. Bis-Tris does not have this issue. Additionally, Bis-Tris demonstrates good buffering capacity and stability within the physiological pH range of approximately 6.8-8.2.
Applications:
- SDS-PAGE: commonly used in SDS-PAGE for separating proteins based on their molecular weight. They provide high-resolution separation of proteins, particularly when used with MOPS or MES running buffers.
- Native PAGE: utilized in native PAGE to study protein complexes in their native state, preserving protein activity and interactions.
- Western Blotting: are used in Western blotting to transfer proteins from gels to membranes. This is crucial for subsequent detection and analysis of specific proteins using antibodies.
- Anion Exchange Chromatography: employed in anion exchange chromatography to maintain a stable pH environment, which is essential for the effective separation of anionic species.
Advantages:
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Bis-Tris is a zwitterionic buffer with good buffering capacity in the pH range of 5.8-7.2, making it suitable for various applications involving proteins and nucleic acids.
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Bis-Tris has minimal interactions with most biomolecules, except for strong binding to Pb2+ and Cu2+ ions.
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High Resolution and Sensitivity: provide sharper band resolution and higher sensitivity compared to traditional Tris-Glycine gels. This is particularly beneficial for detecting low-abundance proteins and for applications requiring high protein integrity, such as post-translational modification analysis and mass spectrometry.
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Versatility: used with different running buffers (MOPS or MES), allowing for the fine-tuning of protein separation based on the size of the proteins. MES is optimal for small proteins (<50 kDa), while MOPS is better for mid-to-large-sized proteins.
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Compatibility with Various Electrophoresis Techniques: compatible with both denaturing and native electrophoresis techniques, making them suitable for a wide range of protein analysis applications.
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Reduced Protein Aggregation: In native PAGE, the neutral pH and compatibility with Coomassie G-250 dye reduce protein aggregation, allowing for better resolution of membrane protein complexes and other sensitive proteins.
Composition
| Ingredients |
Concentration |
| Bicine |
25 mM |
| Bis-Tris |
25 mM |
| EDTA |
1 mM |
Quality Control:
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Appearance: Clear, Colorless Liquid.
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pH: 7.2
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Sterility: Filtered 0.1 micron Twice and 0.04 micron once 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: 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.
