Protease Inhibitor Cocktail with RIPA Buffer
Cat. No. DCP-PICTRIPA1X
Size: 10x10 mL
Sterile; Ultrapure, Filtered 0.1-micron membrane TWICE and 0.04-micron membrane ONCE.
Description:
This Ready-to-use combination of Protease Inhibitor Cocktail with RIPA Lysis Buffer is a powerful tool for efficient cell lysis and protein extraction while preserving protein integrity. RIPA buffer, composed of Tris-HCl, NaCl, detergents (NP-40, sodium deoxycholate, and SDS), and EDTA, effectively disrupts cell membranes and solubilizes proteins. The addition of the protease inhibitor cocktail provides broad-spectrum protection against various classes of proteases, including serine, cysteine, and aspartic proteases. This combination ensures that proteins remain intact during the lysis process and subsequent experimental procedures. Researchers commonly use this mixture for applications such as Western blotting, immunoprecipitation, and various protein assays. The synergy between RIPA buffer's robust lysis capabilities and the protease inhibitor cocktail's protective effects results in high-quality protein extracts, essential for accurate and reliable biochemical analyses. This method is particularly valuable when working with sensitive or easily degraded proteins, as it minimizes proteolysis and maintains the native state of cellular proteins.
The cocktail contains AEBSF.HCl, a serine protease inhibitor; Aprotinin, which targets serine proteases and has anti-inflammatory properties; Bestatin, an aminopeptidase inhibitor; E-64, a cysteine protease inhibitor; EDTA, a metalloproteinase inhibitor; Leupeptin, which inhibits both serine and cysteine proteases; and Pepstatin A, an aspartic protease inhibitor. This cocktail effectively minimizes protein degradation in a wide range of experimental procedures, including cell lysis, protein purification, and enzymatic assays. The carefully balanced concentrations of each component ensure optimal inhibition of proteolytic activity while maintaining compatibility with most downstream applications, making it an invaluable tool for researchers working with sensitive protein samples.
Advantages:
- Prevents protein degradation by inhibiting endogenous proteases released during cell lysis.
- Maintains the integrity and functionality of native cellular proteins.
- Improves the yield of intact proteins for downstream applications.
- Effective for extracting cytoplasmic, membrane, and nuclear proteins.
Application:
- Western Blotting: RIPA buffer is ideal for preparing lysates for Western blot analysis. It efficiently lyses cells and solubilizes proteins, which are then separated by SDS-PAGE and detected using specific antibodies.
- Immunoprecipitation (IP) and Co-Immunoprecipitation (Co-IP): The buffer is effective for IP and Co-IP applications due to its ability to minimize non-specific protein interactions while preserving specific protein-protein interactions.
- Protein Assays: RIPA buffer is compatible with various protein assays, allowing for accurate quantification of protein concentration in cell lysates.
- Reporter Assays: It can be used in reporter assays to study gene expression by measuring reporter enzyme activity in cell extracts.
- Protein Purification: RIPA buffer aids in the extraction and purification of cytoplasmic, membrane, and nuclear proteins from cells, making it suitable for downstream purification processes.
Composition:
| Ingredients |
Concentration |
| AEBSF.HCl |
100 uM |
| Aprotinin |
0.8 uM |
| Bestatin |
0.05 mM |
| E-64 |
1 uM |
| EDTA |
5 mM |
| Leupeptin |
0.01 mM |
| Pepstatin A |
1 uM |
| NP-40 |
1% |
| Tris-HCl |
50 mM |
| Sodium Chloride |
150 mM |
| Sodium deoxycholate |
0.5% |
| SDS |
0.1% |
Quality Control:
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Appearance: Clear solution.
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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 was used for 18 hours 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: -80 ℃
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Shelf life: 3 months
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.
