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AdiFectin™ (CL347)

AdiFectin™ (CL347) Unit size Cat. code Docs Qty Price
TLR7 ligand &nucleic acid carrier
500 µg
tlrl-c347
+-
$157.00

TLR7 Ligand & Nucleic Acid Carrier

AdiFectin™ (bis (phytanyl) N4-{N1-[(4-((6-amino-2-(butylamino)-8-hydroxy- 9H-purin- 9-yl) methyl) benzoyl) glycinyl] sperminyl} propyl phosphonate) is derived from CL307 by conjugation with a bis(phytanyl) phosphonate group.

Addition of this lipid confers to the molecule the ability to form positively charged liposomes, which can encapsulate DNA (or RNA). AdiFectin™ is a weaker TLR7 agonist than CL307, but in contrast to CL307, is able to efficiently complex nucleic acids resulting in a strong IFN response and transgene expression when the nucleic acid is a plasmid DNA carrying an expression cassette.

Repeated in vivo studies have showed that pDNA/AdiFectin™ complexes display robust anti-tumor activity . Tumor growth was markedly reduced resulting in a 50% survival rate. Notably, mice that achieved long-term clearance of tumor following AdiFectin™ treatment were protected from subsequent tumor rechallenge suggesting the generation of a tumor-specific memory immune response.

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Specifications

Specificity: TLR7 agonist and nucleic acid carrier

Synonym: Bis (phytanyl) N4-{N1-[(4-((6-amino-2-(butylamino)-8-hydroxy- 9H-purin- 9-yl) methyl) benzoyl) glycinyl] sperminyl} propyl phosphonate

Working concentration: 300 ng- 3 µg/ml (~200 nM - 2 µM)

Solubility: Ethanol (5 mg/ml)

Formula: C72H134N11O6P

Molecular weight: 1280 g/mol

Endotoxin level: <0.001 EU/µg

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Contents

  • 500 µg AdiFectin™ (CL347)
  • 1.5 ml sterile endotoxin-free water

room temperature Products are shipped at room temperature.

store Stored at -20°C.

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Description

Adifectin (CL347)

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Details

HEK-Blue™ hTLR7 cells stimulation

HEK-Blue™ hTLR7 cells, which stably express an NF-κB-inducible SEAP reporter gene and human TLR7 were incubated in HEK-Blue™ Detection (a SEAP detection growth medium) and stimulated with increasing concentrations of the agonists indicated in the graph.
After 24h incubation, the levels of NF-κB-induced SEAP were determined by reading the OD at 655 nm.

RAW-Blue™ cells, which stably express an NF-kB-inducible SEAP reporter gene, were stimulated with 0.6 mg/ml of multi-PRR ligands

RAW-Blue™ cells, which stably express an NF-κB-inducible SEAP reporter gene, were stimulated with 0.6 µg/ml of InvivoGen’s multi-PRR ligands complexed with 0.1 µg/ml HSV-60 (synthetic dsDNA).
After 24h incubation, the levels of NF-κB-induced SEAP were determined using QUANTI-Blue™, a SEAP detection reagent.

Stimulation of RAW-Lucia cells

RAW-Lucia cells, which stably express an IRF-inducible Lucia, a secreted luciferase reporter gene, were stimulated with 6 µg/ml of InvivoGen’s multi-PRR ligands complexed with 1 µg/ml HSV-60 (synthetic dsDNA). After 24h incubation, the levels of IRF-induced Lucia luciferase were determined using QUANTI-Luc™, a luciferase detection reagent.

Transfection efficiency of AdiFectin™

Transfection efficiency of AdiFectin™.
B16-F1 cells were incubated with pDNAGFP/CL347 complexes at a 1/6 ratio (w/w).
After 48h incubation, GFP expression was detected using fluorescence microscopy. Similar results were obtained with pDNA-GFP complexed with CL419 or CL553. No fluorescence was observed with pDNA-GFP mixed with other multi-PRR ligands, such as CL307 or CL531.

Antitumor effect of CL419, CL347 & CL553

(A) Tumor growth after treatment with CL419, CL347 or CL401complexed with plasmid DNA (pDNA). Seven days after C57BL/6 mice were inoculated subcutaneously with 5.10e5 B16-F1 cells, pDNA/CL419, pDNA/CL347 or pDNA/CL553 complexes were injected intratumorally at a 10:40 (w:w) ratio (10 µg:40 µg/mouse/100 µl) on days 7 and 16. A fourth group received intratumoral injections of the vehicle.

(B) Survival curves for untreated as well as pDNA/CL419-, pDNA/CL347- or pDNA/CL553-treated mice.
Each group contained 8 mice. Black arrows represent the days of injection. Tumor growth was monitored and measured with calipers after day 5 of grafting tumor cells into mice and then every 2 days thereafter. Tumor volume in mm3 was determined according to the formula V = W2 x L/2, where L = length (mm) and W = width (mm).

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