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PamadiFectin™ (CL553)

PamadiFectin™ (CL553) Unit size Cat. code Docs Qty Price
TLR2 & TLR7 Ligand
500 µg
tlrl-c553
+-
$157.00
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TLR2 / TLR7 Ligand & Nucleic Acid Carrier

PamadiFectin™ (N4-(S-((2,3-bis(palmitoyloxy))- (2RS)propyl)-(R)-cysteinyl) N1-(4-(((6-amino-2-(butylamino)-8- hydroxy-9H-purin-9-yl)methyl)benzoyl) glycinyl) spermine) was generated by conjugation of CL307 to a Pam2C group.

PamadiFectin™ induces NF-κB activation through stimulation of both TLR2 and TLR7. In addition, at physiological pH, PamadiFectin™ is able to form complexes with nucleic acids and carry them in the cytosol and nucleus leading to a strong induction of the IRF pathway.
Intratumoral administration of pDNA/PamadiFectin™ complexes leads to spectacular reduction of tumor growth and improved long-term survival in B16-F1 tumor-bearing mice . Tumor rechallenge experiments have not yet been performed.

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Specifications

Specificity: TLR2 / TLR7 agonist and nucleic acid carrier
Synonym:
N4-(S-((2,3-bis(palmitoyloxy))- (2RS)propyl)- (R)- cysteinyl) N1- (4- ( ( (6- amino-2- (butyl amino) -8-hydroxy-9H-purin-9- yl) methyl) benzoyl) glycinyl) spermine
Working concentration:
100 ng- 1 µg/ml (~100 nM - 1 µM)
Solubility:
Water (1 mg/ml)
Formula:
C67H118N12O8S
Molecular weight:
1251 g/mol
Endotoxin level: <0.001 EU/µg

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Contents

- 500 µg PamadiFectin™ (CL553)
- 1.5 ml sterile endotoxin-free water

Products are shipped at room temperature and should be stored at -20°C.

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Description

Pamadifectin structure

PamadiFectin™ (CL553)

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Details

HEK-Blue™ hTLR2 cells stimulation

HEK-Blue™ hTLR2 cells  which stably express an NF-κB-inducible SEAP reporter gene and human TLR2 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.

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 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.

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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