N-(Propargyl-PEG2)-N-Boc-PEG3-t-butyl ester is a click chemistry branched linker. The propargyl group can react with azide-bearing compounds through Click Chemistry, the t-butyl protected carboxyl group can also be deprotected under mild acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
N-(Propargyl-PEG2)-N-Boc-PEG3-t-butyl ester is a click chemistry branched linker. The propargyl group can react with azide-bearing compounds through Click Chemistry, the t-butyl protected carboxyl group can also be deprotected under mild acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Recombinant adeno-associated virus (AAV) vectors are highly promising tools for both in vitro and in vivo gene transfer. Norgen’s AAV Purification Kits provide fast and simple procedures for concentrating and purifying AAV vectors from cell lysate and cell culture media. Purification is based on precipitation onto Norgen Biotek’s proprietary resin. Contaminating cellular debris is largely removed from the sample via a centrifugation step, while contaminating DNA and RNA is reduced using enzymatic digestion. AAV vector purified in this manner is highly active for use in in vitro and in vivo transduction experiments.
AAV Purification Kit
Norgen’s AAV Purification Kit contains sufficient materials for 15 preparations (33.5 mL per prep of supernatant (SN) or a total of 500 mL of supernatant input). Approximately 1 mL of cell pellet can be purified per prep, up to a maximum of 15 mL of cell pellet in total for the entire kit. Up to 33X sample concentration.
AAV Purification Mini Kit
Each spin column is able to concentrate and purify AAV from 0.5-8 mL of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (200 µL). Preparation time for 4 samples is 1.5 hours, with 45 minutes of hands-on time.
AAV Purification Midi Kit
Each spin column is able to concentrate and purify AAV from 8 mL up to 45 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1 mL). The kit may be used to purify up to 8 x 25 mL or 4 x 45 mL of samples using the included columns. Preparation time for 4 samples is approximately 2 to 2.5 hours, with 1.5 hours of hands on time.
AAV Purification Maxi Kit (Slurry Format)
Each spin column is able to concentrate and purify AAV from 45 mL to 90 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 200X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1-10 mL) using the optional concentration step. The kit may be used to purify up to 1 x 900 mL samples or 10 x 45-90 mL samples using the included columns. Preparation time for 1 x 900 mL sample is approximately 2.5 to 3.5 hours, with an optional concentration step requiring an additional 30 min.
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| Kit Specifications | |
| Column Binding Capacity | At least 1 x 1010 AAV particles as determined by qPCR |
| AAV Vector Serotype | Any |
| Average Recovery | > 80% |
| Input Type | Cells, media, or mixed |
| Input Volume | 8 mL – 45 mL |
| Minimum Elution Volume | 1 mL |
| Time to Complete 4 Purifications | 2 – 2.5 hours |
Storage Conditions and Product Stability
DNAse I and RNAse A should be stored at -20°C upon arrival. Elution Buffer O should be stored tightly capped at 4°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. Once opened, the solutions should be stored at 4°C. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 66100 (15 preps) | Cat. 63200 (20 preps) | Cat. 63300 (4-8 preps) | Cat. 63250 (1-10 preps) |
|---|---|---|---|---|
| Lysis Buffer S | 5.5 mL | 5.5 mL | 5.5 mL | 20 mL |
| DNAse I | – | 2 x 25 uL | 2 x 25 uL | 210 μL |
| RNAse A | – | 60 μL | 60 μL | 240 μL |
| HL-SAN Nuclease | 102 μL | – | – | – |
| Binding Buffer A | 20 mL | 4 mL | 4 mL | 2 x 8 mL |
| Purification Solution C | 60 mL | – | – | – |
| Purification Solution D | 130 mL | – | – | – |
| Wash Solution C | 2 x 130 mL | 60 mL | 60 mL | 3 x 60 mL |
| Slurry E | 12.5 mL | – | – | 2 x 14.5 mL |
| Elution Buffer O | 66 mL | 8.5 mL | 8.5 mL | 66 mL |
| Protein Neutralizer | 4 mL | 4 mL | 4 mL | 4 mL |
| Spin Columns | – | 20 | – | – |
| Mini Spin Columns | – | 20 | – | – |
| Midi Spin Columns (grey contents) with Collection Tubes | – | – | 8 | 10 |
| Midi Spin Columns (white contents) with Collection Tubes | – | – | 8 | – |
| Maxi Spin Columns (grey contents) with Collection Tubes | – | – | – | 10 |
| Maxi Spin Columns (white contents) with Collection Tubes | – | – | – | 10 |
| Collection Tubes | – | 40 | – | – |
| Elution tubes (1.7 mL) | 50 | 20 | – | – |
| Midi Elution tubes (15 mL) | – | – | 8 | 10 |
| Maxi Elution tubes (50 mL) | – | – | – | 10 |
| Product Insert | 1 | 1 | 1 | 1 |
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 | 100 mL |
| Yield from 100 mL of Culture | Up to 12 mg |
| Minimum Elution Volume | 4 mL |
| Time to Process 1 Sample | 1 hour |
Storage Conditions
The Cell Lysis Reagent and IB Solubilization Reagent should be stored at 4°C upon receipt of this kit. 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 Binding Buffer C and Binding Buffer N. 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 |
Specification
| 6.5-65 x continuous zoom stereo microscope | XTL-6565A | XTL-6565B | XTL-6565D | XTL-6565C | |
| Zoom Body | Binocular head,45°decline,360°rotate | ● | ● | ||
| Zoom Body | Trinocular head,45°decline,360°rotate | ● | ● | ||
| Eyepiece | WF10X22mm | ● | ● | ● | ● |
| Objective | zoom objective 0.65X—6.5X | ● | ● | ● | ● |
| Zoom Rate | 1:10 | ● | ● | ● | ● |
| Total Amplific- ation Rate | the standard configuration is 6.5X-65X | ● | ● | ● | ● |
| Auxiliary Lens | 0.3X,0.5X,0.7X,1X,1.5X,2X For Option | O | O | O | O |
| WorkingDistance | 110mm | ● | ● | ● | ● |
| Djopte | 55mm-75mm | ● | ● | ● | ● |
| Focus arm | 76mm Diameter,32mm pole size | ● | ● | ● | ● |
| plate | Black/White plate,Glass plate for option | ● | ● | ● | ● |
| Dptiona | Stand | O | O | ||
| Stand | BL3 base,column support upper and lower light source 3 WLED | ● | ● | ||
| Stand | 811 base column support large flat base has no light source | ● | ● | ||
| External light source | LED illumination | ○ | O | O | O |
| Optional | Photography,camera accessories CCD Coupler | 〇 | Q | Q | o |
| Optiona | Display attachment Monitor | O | O | O | |
