SDS Stripping Buffer
Cat No.: DCP-SDSSB1X
Size: 500 mL
Sterile, Ultrapure, Filtered 0.1-micron membrane TWICE and 0.04-micron membrane ONCE.
Description:
In the realm of Western blotting and immunodetection, the choice of stripping buffer plays a crucial role in experimental design and data interpretation. Harsh Stripping Buffer represents a potent tool in the researcher's arsenal, offering a more aggressive approach to antibody dissociation and immunocomplex disruption when conventional mild or medium stripping methods prove insufficient. This buffer's efficacy stems from its unique composition, primarily featuring a high concentration of sodium dodecyl sulfate (SDS) and tris(2-carboxyethyl)phosphine (TCEP). SDS, a powerful anionic detergent, effectively denatures and solubilizes bound antibodies on the membrane. Concurrently, TCEP, a strong reducing agent, cleaves disulfide bonds within the antibody structure and the antibody-antigen complex, further facilitating their dissociation.
The slightly acidic nature of the Harsh Stripping Buffer contributes to its effectiveness by altering charge interactions, thereby disrupting the antibody-antigen binding. This multi-faceted approach ensures a more thorough removal of high-affinity antibodies, which is particularly valuable when re-probing membranes with new antibodies or antigens. It is important to note that the use of Harsh Stripping Buffer may result in some loss of immobilized proteins. However, this potential drawback is often outweighed by the benefit of obtaining cleaner, more reliable signals in subsequent probing steps. This trade-off is especially relevant in scenarios where residual antibodies from previous probing cycles could generate background noise or false-positive results, compromising data integrity.
Advantages:
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It has the ability to remove high-affinity antibodies from the membrane.
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High SDS concentration, reducing agents, elevated temperature, and slightly acidic pH enable it to aggressively denature and dissociate the bound antibodies from the Western blot membrane and immobilized immunocomplex.
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Tris(2-carboxyethyl)phosphine (TCEP) is superior and more effective than dithiothreitol (DTT) or beta-mercaptoethanol to break disulfide bonds.
Composition:
| Ingredients |
Concentration |
| Tris-HCl |
0.5 M |
| SDS |
10% |
| TCEP |
50 mM |
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 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:
CUSTOMIZATION: Please INQUIRE if other concentrations, additions of chemicals/compounds/proteins/supplements, different pH, and modifications are needed.
MILD or MEDIUM stripping buffers with low/high pH and high salt can be customized.
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
