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 | |
| Resin Binding Capacity (total per kit) | At least 5 x 1010 AAV particles as determined by qPCR |
| AAV Vector Serotype | AAV6, AAV9 and others |
| Input Type | Cells, media |
| Input Volume (AAV supernatant) | 1 – 33.5 mL SN per prep (500 mL SN in total) |
| Input Volume (AAV cell pellet) | 1 mL cell pellet per prep (15 mL in total) |
| Time to Complete Purifications | 2.5 to 4.5 hours with 1 hour hands on time |
| In vivo transduction | Yes |
Storage Conditions and Product Stability
HL-SAN Nuclease 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 2 years 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 |
DBCO-NHCO-PEG13-acid is an analog of DBCO-Acid with a hydrophilic PEG spacer arm, which improves water solubility. This reagent is a non-activated building block with enhanced solubility in aqueous media used to derivatize amine-containing molecule through a stable amide bond. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
DBCO-NHCO-PEG13-acid is an analog of DBCO-Acid with a hydrophilic PEG spacer arm, which improves water solubility. This reagent is a non-activated building block with enhanced solubility in aqueous media used to derivatize amine-containing molecule through a stable amide bond. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
The Cylindrospermopsin plate kit is a competitive enzyme-labeled immunoassay. The Cylindrospermopsin sample extract and calibrators are pipetted into the test wells followed by the Cylindrospermopsin antibody into the test wells to initiate the reaction. During the 30 minutes incubation period, Cylin-drospermopsin from the sample and Cylindrospermopsin antigen compete for binding to the Cylindrosper-mopsin antibody. The Cylindrospermopsin antibody is captured on the walls of the test well. Following this 30-minute incubation, the contents of the wells are removed and the wells are washed to remove any unbound Cylindrospermopsin and free Cylindrospermopsin antibody. After wash, 1X HRP-conjugated Antibody#2 is added for 30 minutes incubation. The wells are washed afterwards, and a clear substrate is then added to the wells and any bound enzyme conjugate causes the conversion to a blue color. Following a 15-minute incubation, the reaction is stopped and the amount of color in each well is read. The color of the unknown samples is compared to the color of the calibrators and the Cylindrospermopsin concentration of the samples is derived.
Format:
EPA 10-Day drinking water Health Advisories for Cylindrospermopsin:
Do not Drink – 0.7 μg/L for bottle fed infants and preschool children, pregnant and nursing woman, elderly immunocompromised and liver conditions.
Do not Drink – 3.0 μg/L for school age children to adults.
Do Not Use – 20 μg/L
EPA Draft Human Health Recreational Ambient Water Quality Criteria to protect human health: 8 μg/L.
Format: 96-well microtiter plate (12 test strips of 8 wells)
Standards: 0 | 0.03 | 0.10 | 0.2 | 2 ppb
Incubation Time: 45 Minutes
Compatible for use with US EPA Method 546
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