LDS Sample Buffer with Reducing Agent [4X]
Cat No.: DCP-LDSSBRA4X
Size: 50 mL & 100 mL
Non-Sterile; Ultrapure, Filtered 0.45-micron membrane ONCE.
Description
LDS (Lithium Dodecyl Sulfate) Sample Buffer with a reducing agent is specifically designed for preparing protein samples for denaturing polyacrylamide gel electrophoresis (PAGE). It is used in SDS-PAGE to separate proteins based on molecular weight. The buffer's denaturing and reducing properties allow proteins to migrate through the gel matrix solely based on size, as they all carry a uniform negative charge. This buffer contains TCEP as a reducing agent which can achieve effective protein denaturation and reduction, facilitating accurate analysis through electrophoresis. TCEP breaks disulfide bonds within and between protein molecules, ensuring complete denaturation. It contains lithium dodecyl sulfate, a reducing agent (e.g., TCEP), glycerol (as a density agent), and Coomassie Brilliant Blue and phenol red (as a tracking dye to monitor electrophoresis progress). Coomassie Brilliant Blue is used for visualizing proteins and phenol red is used as a tracking dye during electrophoresis to monitor the progress of the run.
Applications
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SDS-PAGE: Use the LDS Sample Buffer with TCEP for preparing protein samples for SDS-PAGE, particularly when analyzing proteins where maintaining reduced thiol groups is critical.
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Western Blotting: Prepares protein samples for transfer to membranes in Western blotting applications. The denatured and reduced proteins are more readily transferred and accessible for antibody binding.
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Protein Analysis: Facilitates the analysis of protein composition, purity, and molecular weight. It is particularly useful for identifying and quantifying proteins in complex mixtures.
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Comparative Studies: Allows for the comparison of reduced and non-reduced protein samples, providing insights into the role of disulfide bonds in protein structure and function.
Advantages:
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Uniform Charge: By denaturing and reducing proteins, the buffer ensures that all proteins carry a uniform negative charge. This is crucial for accurate separation based solely on molecular weight during SDS-PAGE.
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Enhanced Protein Solubilization: The presence of LDS, a detergent similar to SDS, helps solubilize proteins effectively. This is particularly important for membrane proteins or those that are otherwise difficult to solubilize.
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Complete Denaturation: The combination of LDS andTCEP, ensures that proteins are fully denatured. This disrupts both non-covalent interactions and disulfide bonds, allowing proteins to unfold completely into linear chains.
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Tracking and Density: Glycerol is included as a density agent to ensure that samples sink into the wells of the gel. Coomassie Brilliant Blue serves as a tracking dye to monitor the progression of electrophoresis.
Composition:
| Ingredients |
Concentration |
| Tris-HCl |
988 mM |
| EDTA |
2.04 mM |
| LDS |
8% |
| Glycerol |
40% |
| Coomassie blue |
0.88% |
| Phenol red |
0.7 mM |
| TCEP |
50 mM |
Quality Control:
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Appearance: Dark Blue Liquid.
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pH: 8~8.4
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Sterility: Filtered 0.45 micron once.
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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: 6 months
CUSTOMIZATION: Please INQUIRE if other concentrations, additions of chemicals/compounds/proteins/supplements, different pH, and modifications are needed.
* 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.
