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in vivo siRNA transfection kit "AteloGene® Local Use Quick Gelation"

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Features and Advantages

Quick gelation at injection sites!

AteloGene Local Use “Quick Gelation” (AteloGene QG) and AteloGene Local Use were mixed with siRNA respectively, and were observed for gelation after administration in mouse subcutaneous tissue. Compared with AteloGene Local Use, results for AteloGene QG showed a rapider gelation after administration.

Efficient Delivery!

AteloGene QG and AteloGene Local Use were mixed with Luciferase siRNA (Luc siRNA) respectively, and evaluated for the luminescence of Luciferase after administration in a Dual-luciferase expressed subcutaneous tumor model. Result with AteloGene QG showed higher inhibition of Luciferase gene expression than results compared to AteloGene Local Use.

Increased Volume! 

Kit component

Contents of kit
This kit is intended for 15 times of administration.

1) Prefilled syringe (filled with “AteloGene® QG”)
Each syringe is for 5 times of administration, including losses during the preparation steps.
540 µL × 3 syringes
2) QG buffer: 2.5 mL × 1 bottle
3) 2 mL microtube:
A spare tube is included. 
4 tubes
4) 18G needle (for ejection and suction):
2 spare needles are included.
8 needles
5) Instruction manual: 1 leaflet

 

Devices and reagents required other than the kit

- 1 mL dispossable syringe
- 27G needle (for injection)
- siRNA/miRNA (PAGE- or HPLC- purified grade)
- Container for preparation of siRNA/miRNA solution (sterilized, RNase free)
- Cooling device (crushed ice, cold block, etc.)
- Tube rotator (that can tumble and agitate such as TAITEC RT-5, Bibby scientific SB3, etc.)
- Pipetter and tips (sterilized, RNase free)
- High-speed refrigerated centrifuge
- Anesthetic

 

Storage
Storage temperature: 2-10°C (do not freeze)
Precautions for storage

- Gelation and thermal denature occur in “AteloGene®” at a temperature higher than 20°C. Do not use “AteloGene®” that was once gelated or thermally denatured.
- Freezing “AteloGene®” causes bubbles in the mixture and the dispersion of components. Never use “AteloGene®” that was once frozen.

 

Precautions for use

1)

Do not use “AteloGene® ” for any purpose other than research use. Application to the human body is strictly prohibited.

2)

Be sure to read the instruction manual before use. The manufacturer is not liable for the results of usage by methods other than that described in the instruction manual. The expected effects may not always be obtained depending on the siRNA/miRNA sequences, administration targets or methods.

Methods

“AteloGene® Local Use Quick Gelation” Preparation Instruction 

In this section, the number 1)-4) refer to the "Kit contents" of the product page. Implement measures to secure an RNase-free environment, as for as possible, to avoid the degradation of siRNA/miRNA in advance.

1)Preparation of “AteloGene® Local Use Quick Gelation”

Attach the 4) 18G needle to the 1) Prefilled syringe. Eject the whole amount (540 µL) into the 3) 2 mL microtube. After ejection, cool the microtube containing “AteloGene® QG” on ice.

Note) An excess amount of “AteloGene® QG” is provided in the 1) Prefilled syringe so that 540 µL of “AteloGene® QG” can be injected into the 3) 2 mL microtube regardless of losses during the preparation steps.

2)Preparation of siRNA solution

Prepare 25-50 μM siRNA/miRNA solutions with RNase-free water.

3)Preparation of "AteloGene®" & siRNA mixed solution.

While cooling on ice, gently pour 540 µL of the 2) QG buffer and then 120 µL of the siRNA/miRNA solution onto 540 µL of “ AteloGene® QG” in the 3)2mL microtube. Rotate and mix the solutions at 4°C for 10 minutes. To avoid forming bubbles, the rotation speed should be approximately 12 r.p.m. (in the case of a rotator with a diameter of 20 cm).

Note) Although the mixture may become cloudy during this procedure, it will become a clear solution as the components are thoroughly mixed by rotation.

4) Deformation of bubbles and preparation for administration

Centrifuge the mixture at 10,000 r.p.m. for 1 minute at 4°C to deform bubbles. Attach the 4)18G needle to the disposable syringe and slowly draw the mixture while avoiding forming bubbles. Replace the needle with the 27G needle and keep the syringe refrigerated until immediatelly before administration.

Mechanism of siRNA transfection using “AteloGene®”.  Mechanism

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Product List

Product Name Cat# Quantity Price

AteloGene Local Use Quick Gelation

KOU-1492 1KIT

¥ 81,300
$ 1084
€ 813

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.