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| Protein Staining and other Solutions 1 » SMART™ Bacterial Protein Extraction Solution (110 |
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| SMART™ Bacterial Protein Extraction Solution (110 |
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The SMART¢â> Bacterial Protein Extraction Solution is designed for fast and easy extraction of total proteins from bacteria without the need for sonication or precipitation. This solution utilizes a proprietary non-ionic detergent in 20mM Tris/HCl(pH7.5). In addition, the SMART¢â Bacterial Protein Extraction Solution offers several folding increase in the yield of soluble protein when compared with other commercial lysis reagent. Depending on the particular application, additional components, such as lysozyme, protease inhibitors, salts, reducing agents and chelating agents may be added to the solution. The solution may be used for both soluble protein extraction and inclusion body purification from bacterial cell lysates. In fact, yields obtained with SMART¢â Bacterial Protein Extraction Solution greatly exceeds those obtained using standard sonication methods!
Bacteria often over-express recombinant proteins and form inclusion bodies, which are insoluble aggregates of misfolded protein. Centrifugation separates inclusion bodies from soluble proteins; however, Lysozyme is required for purification of inclusion bodies. Lysozyme significantly improves inclusion body purity by digesting the cell debris.
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- Non-ionic detergent in a Tris/HCl (pH7.5) buffer. Therefore, use Tris buffer for subsequent protein purification. Eliminates mechanical disruption and provides a gentle and efficient method for extracting proteins.
- Whole cell lysates prepared with the SMART¢â Bacterial Protein Extraction Solution are compatible with the SMART¢â BCA Protein Assay Kit(Cat.No.21071) and coomassie protein assays.
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* SMARTTM Bactrial Protein Extraction Solution
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110 §¢
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* protease inhibitor is not included
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1)
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Upon receipt store product at room temperature. |
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Comparison of Protein Recovery with Competitor |
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Fig. 1. Comparison of efficiency of protein extraction with competitor¡¯s Kit.
The results of Table 1 was demonstrated graphically. The SMART¢â Bacterial Protein Extraction Solution showed improved recovery of bacterial protein extraction. |
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Table. 1. Comparison of extracted protein concentration with competitor
The Quantities of extracted proteins were estimated using by SMART¢â BCA Protein Assay Kit (Cat.No 21071) |
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Inclusion Body Isolation |
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Fig. 4. Isolated inclusion bodies
Each of inclusion body of several expression hosts was isolated and solubilized (followed by Protocol 2).
Lane A, M15 [rGroEL]; lane B, BL21[rTaq ]; lane C, origami [rTaq]; lane D, Rogettagami [rTaq]; lane M, Protein Markerlane 1, SMART¢â Bacterial Protein Extraction Solution; lane 2, Competitor¡¯s Kit |
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