Hypertonic Buffer
Cat. No. DCP-HB1X
Size: 500 mL
Sterile; Ultrapure, Filtered 0.1-micron membrane TWICE and 0.04-micron membrane ONCE.
Description:
Hypertonic buffer is a specialized solution designed for cell extraction, particularly effective for nuclear protein extraction. It contains potassium chloride (KCl), which creates a high-salt environment crucial for extracting proteins from nuclei. This buffer is supplemented with HEPES-KOH, a zwitterionic buffering agent that maintains a stable pH of 7.8, ideal for preserving protein structure and function. The addition of magnesium chloride (MgCl2) provides essential magnesium ions, which act as cofactors for many enzymes and help stabilize nuclear structures. The buffer also includes EDTA, a chelating agent that binds metal ions, thereby inhibiting metal-dependent enzymes that could degrade proteins or nucleic acids. The 20% glycerol content serves as a stabilizing agent, protecting proteins from denaturation during extraction and storage. This carefully formulated buffer is optimized to maintain osmotic balance, preserve protein integrity, and facilitate efficient extraction of nuclear proteins from cells.
Advantages:
- Osmotic effects: The high solute concentration in hypertonic buffers can cause cells to shrink, which can be beneficial for certain extraction procedures, particularly for nuclear proteins.
- pH stability: Hypertonic buffers, especially those containing components like HEPES, maintain a stable pH despite changes in carbon dioxide concentration. This is crucial for maintaining optimal conditions for enzymatic reactions and protein stability.
- Protein stabilization: The high salt concentration in hypertonic buffers can help stabilize proteins by reducing their solubility and preventing denaturation. This is particularly useful in protein purification and storage.
- Enzyme activity modulation: The high ionic strength of hypertonic buffers can influence enzyme activity, often enhancing the function of certain enzymes that require specific salt concentrations for optimal performance.
- Reduced contamination: The high salt concentration in hypertonic buffers can inhibit the growth of many microorganisms, potentially reducing contamination risks in long-term experiments.
- Selective extraction: Hypertonic buffers can be used to selectively extract certain cellular components, such as nuclear proteins, while leaving other structures intact.
Application:
- Nuclear Protein Extraction: This buffer is well-suited for extracting nuclear proteins from cells. The high salt concentration is effective in extracting proteins from nuclei after cell lysis and separation of cytoplasmic and nuclear fractions.
- Cell Lysis: While not a complete lysis buffer on its own, it can be used as part of a cell disruption protocol. The hypertonic nature causes cells to shrink, potentially weakening cell membranes and facilitating subsequent lysis steps.
- Protein Stabilization: The combination of HEPES buffer and glycerol helps maintain protein stability during extraction and purification processes. This is crucial for preserving protein structure and function for downstream applications.
Composition:
| Ingredients |
Concentration |
| HEPES-KOH |
50 mM |
| Potassium Chloride |
0.42 M |
| Magnesium Chloride |
5 mM |
| EDTA |
0.1 mM |
| Glycerol |
20% |
Quality Control:
- Appearance: Clear solution.
- pH: 7.8
- 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 was used for 18 hours 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 other tracking dyes, 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.
