Gene Tools - Model Endo-Porter - Simple Delivery Reagent

Detailed studies with a FITC-anti-actin antibody fragment (Sigma F-3046) and fluorescent high-molecular weight dextrans suggest Endo-Porter can deliver high concentrations of cargo exceeding 70kD into both adherent and non-adherent cells. Unlike the many delivery reagents that claim effectiveness in the presence of serum, Endo-Porter actually works with serum concentrations up to the 10% standard. Endo-Porter is excellent at delivering peptides, proteins, and Morpholino antisense oligos as well! We do not recommend using Endo-Porter for delivery of large (>100 Kd) proteins, as efficacy is poor.

siRNA delivery: Endo-Porter has been reported effective for delivering siRNA in HeLa, Huh-7 and Hek293T cultures [Bartz et al. 2011]. Combined with poly-lysine, Endo-Porter delivered siRNA efficiently in rat hepatoma H4IIEC3 cells [Inoue et al. 2008]. Endo-Porter has been reported to deliver siRNA in vivo when encapsulated in glucan shells [Tesz et al. 2011, Aouadi et al. 2009]. Our tests have not found Endo-Porter effective for delivery of plasmid DNA.

View or download an animation of Endo-Porter`s mechanism of action by selecting one of the following two choices:

Windows Media Endo-Porter animation
Apple QuickTime Endo-Porter animation

Animation by Pathworks of Corvallis, OR.

2. What are the advantages of Endo-Porter over other delivery reagents?
Most other delivery reagents require pre-incubation or optimized reagent-to-cargo ratio in order to achieve delivery, but not Endo-Porter. You can add as much or as little cargo as desired and use the same amount of Endo-Porter EVERY time. Endo-Porter lasts indefinitely and can be stored at room temperature or below.

3. What do I get and how much does it cost?
All Endo-Porter formulations cost $200. We recommend Endo-Porter PEG for all new users, as this is our highest efficacy formulation with even less toxicity than the previous-best DMSO formulation. Our legacy formulations are still available for replicating previous experiments.
The available formulations are:

1. Endo-Porter PEG (preferred), 1 mM Endo-Porter in 10% polyethylene glycol 1500 MW, aqueous.
2. Endo-Porter DMSO (legacy), 1 mM Endo-Porter in DMSO.
3. Endo-Porter Aqueous (legacy), 1 mM Endo-Porter in an aqueous solution of mannitol.

You will receive 1.0 mL of 1.0 mM Endo-Porter in solution. This is enough to deliver up to about 150 mL of cells with Endo-Porter (optimal dose varies with the cell type). Endo-Porter can be ordered online today!

For additional questions or assistance from our Ph.D.-level customer support staff use our online live help, submit a question, or call (541) 929-7840 ext. 1.

4. Endo-Porter excels in independent comparison of nine cytosolic delivery reagents
When delivering Atto488-bovine serum albumin into cultured human sarcoma cells using nine different commercial delivery products, only Endo-Porter achieved clear cytosolic delivery.
"A primarily endosomal uptake of proteins is consistent with the observation that cytoplasmic signals could only be observed in Endo-Porter treated cells. The Endo-Porter reagent is a specialized reagent to disrupt endosomal membranes as a consequence of endosomal acidification and it is the only one of the reagents tested which has exclusively been invented to overcome endosomal entrapment."[1]
Endo-Porter also delivered to the largest fraction of cells: "Comparing the different transduction reagents, the percentage of fluorescence positive cells varied between 89.2 (TurboFect) and 96.8% (Endo-Porter)."[1]
[1] Mellert K, Lamla M, Scheffzek K, Wittig R, Kaufmann D. Enhancing endosomal escape of transduced proteins by photochemical internalisation. PLoS One. 2012;7(12):e52473. doi: 10.1371/journal.pone.0052473. Epub 2012 Dec 21.

5. The protocol is simple: squirt and swirl.
Our recommended delivery procedure:

1. Replace spent culture medium with fresh medium (preferably 10% serum).
2. Add the desired cargo and swirl to mix.
3. Add 6 µL of Endo-Porter for every 1 mL of culture medium & cargo and immediately swirl to mix.

In most cases, long-term (72 hour) and short-term (24 hour) delivery is optimal at 6 µM Endo-Porter (6 µL/mL); however, cell lines vary in their response to Endo-Porter and it is prudent to start experiments with a new cell line by doing a ranging study for efficacy and toxicity at 2, 4, and 6 µL/mL Endo-Porter. Check the cells at 72 hours for signs of toxicity, then use the highest concentration that the cells tolerate for subsequent delivery procedures. Endo-Porter PEG may be tolerated at even higher doses. Unlike most delivery solutions, 10% serum concentrations yield better results than reduced serum levels. Additional cargo and Endo-Porter can be added with each media replacement if desired. For delivery of Morpholino oligos, we suggest you start with 10 µM oligo final concentration in the culture medium then adjust concentrations as needed to achieve effective and specific gene knockdown.