Oligo(deoxyadenosine:deoxythymidine) (endotoxin-free) (synthetic)

Oligo(dA:dT)

IFN

Activator

<0.0002EU/µg (LAL test; BioWhittaker)

Lyophilized. Sterile.

Aliquoted by weight of dry material. Contains polymer, residual salt and water. Sufficient for at least 200 cellular activation assays at 0.01-0.025µg/ml.

For a 250µg/ml stock solution, dissolve the total vial content in 200µl endotoxin-free buffer or water.

Synthetic.

For in vitro stimulation we recommend the following procedure:
1. Bone marrow-derived mouse dendritic cells (BM mDC) are cultured 8×10⁴/100μl in RPMI containing 10% FCS over night at 5% CO₂ and 37°C.
2. The medium is removed and replaced by 100μl OPTI MEM (Invitrogen) and subsequently the transfection mixture is added: 25μl OPTI MEM containing 0.025 to 0.25 Poly(dA:dT) + 25μl OPTI MEM containing 0.5μl Lipofectamine 2000 (Invitrogen) or alternative transfection agents.
3. Over night stimulation at 5% CO₂ and 37°C.
4. Measurement of IFN-α/β in culture supernatants.