These kits provide everything required to isolate and purify inclusion body proteins from induced bacterial cultures. First a proprietary Cell Lysis Reagent is used to selectively lyse the cells and release inclusion bodies in their solid form. Next, inclusion bodies are dissolved and their contents released using the provided IB Solubilization Reagent. Inclusion body proteins are then further purified using spin columns for rapid and convenient buffer exchange and desalting. This kit provides a convenient way to screen recombinants prior to scaling up.
ProteoSpin™ Inclusion Body Protein Isolation Micro Kit
The process is efficient and streamlined and can process up to 12 samples in only 60 minutes. Each spin column is able to recover up to 50 µg of acidic or basic proteins. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.
ProteoSpin™ Inclusion Body Protein Isolation Maxi Kit
The procedure is efficient and streamlined and can process up to 4 samples in approximately 2 hours. Each spin column is able to recover up to 12 mg of acidic or basic proteins from 100 mL of induced bacterial culture. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.
About Inclusion Bodies
Bacteria are widely used for the expression of different proteins. However, 70-80% of the proteins expressed in bacteria by recombinant techniques are typically contained in insoluble inclusion bodies (i.e., protein aggregates). The protein of interest found in these subcellular structures is often inactive, due to incorrect folding. The production rate of recombinant proteins stored in inclusion bodies is invariably higher than those synthesized as soluble proteins. The reason behind this is thought to be the resistance of insoluble proteins to proteolysis by cellular enzymes. In addition, separation of insoluble recombinant proteins in inclusion bodies is considerably easier than that of soluble proteins. These factors have been the major influences favoring scale-up of high-value proteins using bacterial fermentation for example. Procedures for the purification of the expressed proteins from inclusion bodies are often labour-intensive, time-consuming and not cost-effective. This kit provides the essential reagents for cell disruption, inclusion body solubilization and purification using spin column chromatography – all optimized to work together thereby simplifying the process and saving a tremendous amount of time and cost.
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| Kit Specifications | |
| Maximum Culture Volume | 1.5 mL |
| Yield from 1.5 mL | Up to 50 μg |
| Minimum Elution Volume | 30 μL |
| Time to Process 12 Samples | 60 minutes |
Storage Conditions
The Cell Lysis Reagent and IB Solubilization Reagent should be stored at 4°C upon receipt of this kit. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. Once opened, the solutions should be stored at 4°C when not in use except for the pH Binding Buffers (Acidic and Basic). Some precipitation will occur with 4°C storage. This precipitation should be dissolved with slight heating to room temperature before using.
| Component | Cat. 10300 (Micro – 25 preps) | Cat. 17700 (Maxi – 4 preps) |
|---|---|---|
| Wash Solution C | 30 mL | 130 mL |
| Wash Solution N | 30 mL | 130 mL |
| Binding BUffer A | 4 mL | 20 mL |
| Binding Buffer N | 4 mL | 20 mL |
| Elution Buffer C | 8 mL | 2 x 30 mL |
| Protein Neutralizer | 4 mL | 4 mL |
| Cell Lysis Reagent | 15 mL | 110 mL |
| IB Solubilization Reagent | 2 mL | 50 mL |
| Syringes, 1cc, slip tip | 25 | – |
| Needles (Bev, 20G x 1 inch) | 25 | – |
| Syringes, 10 mL, Luer-Lok™ Tip | – | 4 |
| Needles (18G x 1.5 inch) | – | 4 |
| Micro Spin Columns | 25 | – |
| Maxi Spin Columns (filled with SiC) inserted into 50 mL collection tubes | – | 4 |
| Collection Tubes | 25 | – |
| Elution Tubes (1.7 mL) | 25 | – |
| Elution Tubes (50 mL) | – | 4 |
| Product Insert | 1 | 1 |
Tri-(Propargyl-PEG2-ethoxymethyl)-methane-amido-PEG3-carboxylate is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The terminal carboxylic acid can react with primary amino groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Tri-(Propargyl-PEG2-ethoxymethyl)-methane-amido-PEG3-carboxylate is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The terminal carboxylic acid can react with primary amino groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Usages:
Beef extract powder is a purified extract of beef .Used with meat and casein peptones as a source of nutrients in culture media .
Technical specification:
Total nitrogen(TN)—————————≥12.0%
Amino nitrogen(AN) ————————≤2.7%
Chlorides————————————–＜6%
Moisture—————————————≤5.0%
PH———————————————–6-7
Storage: Keep container tightly closed, store in a cool, dry place, away from bright light.
Specifications: 500g/bottle; 10kg /bag
500g/bottle;10KG/bag