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New type Protein Transfection Reagent ProteoCarry | Funakoshi

Date:November 28 2017Web Page No:80968

Funakoshi Co.,Ltd.

Going to Cytoplasm, not ending up in endosome!
ProteoCarry

ProteoCarry is a new type peptide base transfection reagent.

When using a conventional protein transfection reagent, transfected proteins are accumulated in endosome or lysosome, and most of such proteins are dissociated in lysosome proteolysis system.
Contrary, ProteoCarry can deliver transfected proteins to cytoplasm effectively.
Moreover, pre-incubation with proteins are NOT needed.
Therefore effect to the protein functions can be minimized.

Product Background (click here)

ProteoCarry is a novel peptide-based transfection reagent for exogenous proteins such as functional proteins, peptides and antibodies with highly efficiency of cytosolic delivery. Protein transfection, intracellular delivery of proteins, is a powerful method to analyze cellular response to the protein of interest. While plasmid-based gene expression generally needs 12-24 hours to express proteins well, protein transfection reagents can immediately import functional proteins into living cells within several hours. Although many protein transfection reagents including peptide-, cationic lipid- and polymer-based compounds have been developed to date, these reagents struggle to deliver proteins into “cytosol”. Commonly these reagents interact with protein to form complexes and subsequently protein-reagent complexes are entered into cells via endocytosis pathway. But protein-reagent complexes are able to escape endosomes with little efficiency and consequently transported to lysosomes to be degraded. Highly effective endosomal escape of proteins is the most important subject of protein transfection reagents. ProteoCarry can overcome this problem based on a novel and potent pH-dependent endosomal membrane-lytic activity. Proteins and ProteoCarry can be delivered into endosomes by endocytosis pathway and also escape to cytosol by its highly membrane-lytic activity with low cytotoxicity.

About protein transfection reagent and issue of conventional one

Features

  • Highly soluble in water
  • High efficiency of delivering proteins to cytoplasm
  • Fast - 1 hour incubation
  • No pre-incubation with proteins needed
  • Effect to the function of proteins cab be minimized
  • Low cytotoxicity
  • Polysaccharide can be also transfected.

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Procedure overview

Procedure overview

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Kit components

  • ProteoCarry, 4mg in vial (vial A)
  • FITC-dextran as positive control, 2 mg in vial (vial B)

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Application Examples

  • Intracellular delivery of exogenous or recombinant proteins including peptides, enzymes, antibodies
  • Intracellular delivery of bio-macromolecules including glycosaminoglycans etc.

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Storage

  • Shipping : Room temperature
  • Store : -20℃

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Validated cell types

  • HeLa, SW280, COS7, NIH3T3, HUVEC

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Application Data

Transfection of fluorophore-conjugated dextran

Transfection of fluorophore-conjugated dextran
HeLa cells were treated with green fluorophore-conjugated dextran (Dextran-FL, average MW 10 kDa, final 200 μg/ml) in the presence or absence of ProteoCarry (1x conc.) for 1 hour at 37oC. Without ProteoCarry, fluorescent signals were detected from endosome-like dot structures inside cells. On the other hand, ProteoCarry clearly stimulated cytosolic delivery of Dextran-FL.

Transfection of fluorophore-conjugated IgG

Transfection of fluorophore-conjugated IgG
HeLa cells were treated with green fluorophore-conjugated IgG (IgG-FL, final 50 or 500 μg/ml) in the presence or absence of ProteoCarry (1x conc.) for 1 hour at 37oC. Without ProteoCarry, IgG-FLs were aggregated into endosome-like dot structures. On the other hand, ProteoCarry promoted cytosolic delivery of IgG-FL even if some IgG-FLs were detected in endosomes.

Transfection of fluorophore-conjugated anti-nucleus pore complex (NPC)

Transfection of fluorophore-conjugated anti-nucleus pore complex (NPC)
HeLa cells were treated with red fluorophore-conjugated anti-nucleus pore complex (NPC) antibody (anti-NPC-FL, final 50 μg/ml) in the presence or absence of ProteoCarry (1x conc.) for 1 hour at 37oC. Without ProteoCarry, a little of fluorescent signal was observed in dot-like structure. On the other hand, ProteoCarry clearly promoted localization of anti-NPC-FL to peri-nuclear structure. This result indicates that ProteoCarry could induces incorporation of anti-NPC-FL into cytosol and binding of antibody to nucleus pore complex.

Transfection of nuclear-localizable EGFP

Transfection of nuclear-localizable EGFP
HeLa cells were treated with nuclear localization signal (NLS)-tagged EGFP (NLS-EGFP, final 10 μM) in the presence or absence of ProteoCarry (1x conc.) for 1 hour at 37oC. Without ProteoCarry, a little of fluorescent signal was observed in endosome-like dot structures. On the other hand, ProteoCarry promoted nuclear-localization of NLS-EGFP.

Transfection of Cre recombinase for Cre/lopX recombination assay

Transfection of Cre recombinase for Cre/lopX recombination assay
HeLa cells were transfected with a plasmid which encodes loxP-DsRed-stop-loxP-EGFP sequence by conventional lipofection reagent. Next day, the HeLa cells were treated with recombinant Cre recombinase (final 10 μM) for 1 hour at 37oC with or without ProteoCarry (1x conc.). Without ProteoCarry, almost cells still keep DsRed expression under the treatment of Cre recombinase. On the other hand, ProteoCarry promoted conversion from DsRed to EGFP by Cre/loxP recombination.

Transfection of Saporin enzyme

Transfection of Saporin enzyme
Saporin is an enzymatically cytotoxic protein which binds to ribosomes and inactivates ribosomes via its RNA glycosidase activity. RAW264.7 cells were treated with saporin (final 10 μg/ml) with or without ProteoCarry for 1 hour at 37oC. After 24 hours culture, cell viability was measured by MTT assay. Without ProteoCarry, saporin induced cell death with less than 30% efficiency. On the other hand, ProteoCarry dramatically increased cytotoxicity of saporin and almost cells were dead (>90%).

Reference data: Intercellular localization of FL-labeled ProteoCarry

Reference data: Intercellular localization of FL-labeled ProteoCarry
To monitor intracellular localization of ProteoCarry, fluorophore-labeled ProteoCarry (ProteoCarry-FL) was prepared and HeLa cells were treated with ProteoCarry-FL. Fluorescent signal was observed by confocal microscopy. Signal was detected mainly in endosomal dot like structure and little fluorescent signal was observed in cytosol. This data indicates ProteoCarry keeps binding to endosomal membrane after lysis of endosomes.


[Date : February 27 2021 00:28]

Detail Product Name Product Code Supplier Size Price
ProteoCarry <Protein Transfection Reagent>
DatasheetThis may not be the latest data sheet.
FDV-0015 FNAFunakoshi Co.,Ltd. 1 set $400

Description ProteoCarry is a new type peptide base transfection reagent. When using a conventional protein transfection reagent, transfected proteins are accumulated in endosome or lysosome, and most of such proteins are dissociated in lysosome proteolysis system. Contrary, ProteoCarry can deliver transfected proteins to cytoplasm effectively. Moreover, pre-incubation with proteins are NOT needed. Therefore effect to the protein functions can be minimized.
Storage -20°C CAS
Link

[Date : February 27 2021 00:28]

ProteoCarry <Protein Transfection Reagent>


  • Product Code: FDV-0015
  • Supplier: FNA
  • Size: 1set
  • Price: $400

Description ProteoCarry is a new type peptide base transfection reagent. When using a conventional protein transfection reagent, transfected proteins are accumulated in endosome or lysosome, and most of such proteins are dissociated in lysosome proteolysis system. Contrary, ProteoCarry can deliver transfected proteins to cytoplasm effectively. Moreover, pre-incubation with proteins are NOT needed. Therefore effect to the protein functions can be minimized.
Storage -20°C CAS
Link

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CONTACT

export@funakoshi.co.jp

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