Cytoskeleton Extraction Buffer [1X]
Cat No.: DCP-CEB1X
Size: 100 mL & 500 mL
Sterile, Ultrapure, Filtered 0.1-micron membrane TWICE and 0.04-micron membrane ONCE.
Description
The cytoskeleton-bound protein Extraction Buffer, also called the Cell Pellet extraction buffer, is a key component for selectively isolating and enriching cytoskeletal proteins from cells for further analysis.
Cell lysis buffers such as RIPA buffer are only suitable for extracting nuclear, cytoplasmic and mitochondrial protein. The cytoskeleton-bound protein extraction requires a specialized buffer to solubilize and extract the cytoskeletal proteins that remain insoluble after the typical cell lysis treatment and extraction steps.
It is a detergent-based buffer (50 mM PIPES, 50 mM NaCl, 5% glycerol, 0.1% NP-40, 0.1% Triton X-100 and 0.1% Tween 20) that retains focal adhesion and actin-associated proteins while removing soluble cytoplasmic and nuclear proteins. This allows for the selective enrichment of cytoskeleton-associated proteins. It contains protease and phosphatase inhibitors to prevent degradation and preserve post-translational modifications of the cytoskeletal proteins. After solubilization, the cytoskeletal proteins can be further processed and analyzed using mass spectrometry-based proteomic techniques.
The buffer is used to lyse cells grown in culture dishes. The insoluble cytoskeletal proteins remain attached to the dish while the soluble proteins are collected in the lysate. After washing with PBS, the ice-cold buffer is added to the dish and incubated for 1.5 minutes on ice to allow lysis.
Advantages:
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It allows for more complete and in-depth profiling of the "cytoskeletome" - the full set of cytoskeletal and cytoskeleton-associated proteins.
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It is more efficient at extracting and solubilizing cytoskeletal proteins compared to other protein extraction buffers like RIPA buffer.
Composition:
| Ingredients |
Concentration |
| Tris-HCl |
10 mM |
| NaCl |
100 mM |
| EDTA |
1 mM |
| EGTA |
1 mM |
| NaF |
1 mM |
| Na4P2O7 |
20 mM |
| Na3VO4 |
2 mM |
| Triton X-100 |
1% |
| Glycerol |
10% |
| SDS |
0.1% |
| Deoxycholate |
0.5% |
Quality Control:
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Appearance: Clear, Colorless Liquid.
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pH: 7.4
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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: 12 months
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
