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商品編號


XPAK730PA-1



品牌


SBI



公司


System Biosciences（SBI）



公司分類(lèi)


exosome labeling



商品信息

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Overview:
Label exosomes with GFP using XPack
TM

Easily create stable cell lines that produce GFP-labeled exosomes with the XPack MSCV-XP-GFP-EF1α-Puro Expression Lentivector. Take advantage of
SBI
’s well-proven lentivector technology to express GFP from the strong MSCV promoter for expression in hematopoietic and stem cell lines (also available with a CMV promoter for expression in most cell lines) and the optimized XPack exosome targeting tag to package GFP into exosomes.


The XPack System at a glance:


Powerful—enables a wealth of applications, including

Exosome labeling

Exosome cargo tracking

Exosomes as cargo delivery vehicles

Exosomes as biotherapeutics

Exosome-generating stable cell lines with protein cargo-of-interest



Optimized—
SBI
developed the XPack peptide tag for robust exosome targeting

Reliable—based on
SBI
’s highly-cited, well-proven lentivector technology

Versatile

Two promoter options for expression in a wide range of cells

Comes as pre-built reporters (GFP, RFP, luciferase) or ready-to-customize cloning lentivectors




Choose the XPack Lentivector that’s right for you




?

CMV Promoter
For most cell lines

MSCV Promoter
For hematopoietic and stem cell lines

?

?





Customize with our cloning vector



?

XPAK510PA-1

XPack CMV-XP-MCS-EF1α-Puro

XPAK710PA-1

XPack MSCV-XP-MCS-EF1α-Puro



Label exosomes with pre-built reporters



GFP

XPAK530CL-1
XPAK530PA-1

XPAK530VA-1

XPack CMV-XP-GFP-EF1α-Puro

XPAK730PA-1
XPAK730VA-1

XPack MSCV-XP-GFP-EF1α-Puro



RFP

XPAK531PA-1

XPack CMV-XP-RFP-EF1α-Puro

XPAK731PA-1

XPack MSCV-XP-RFP-EF1α-Puro



Luciferase

XPAK532CL-1
XPAK532PA-1

XPAK532VA-1

XPack CMV-XP-Luciferase-EF1α-Puro

XPAK732PA-1
XPAK732VA-1

XPack MSCV-XP-Luciferase-EF1α-Puro




Supporting Data:

XPack efficiently packages GFP into exosomes which can be used to deliver GFP to target cells


Part 1: Packaging GFP into exosomes with XPack

The XPack-GFP fusion protein is efficiently packaged as exosome cargo and delivered to target cells. HEK-293 cells were transfected with a plasmid encoding an N-terminal XPack-GFP fusion protein (Cat.# XPAK530PA-1). Twenty-four hours later, GFP localization patterns were visualized on a
Leica
DMI300B fluorescent microscope (Figure 1, upper panel). GFP expression patterns within transfected cells demonstrate a localization pattern characteristic of exosomal lo
ADI
ng, with punctate spots along the cell periphery indicating budding vesicles and a brightly fluorescent spot in the cell interior corresponding to the multivesicular body (MVB), the site of exosome assembly and maturation prior to secretion.

After 48 hours, exosomes were isolated using ExoQuick-TC
?
and analyzed for GFP via Western blot (Figure 1, lower panel). Equal amounts of exosomal lysates (15 ?g) were separated on gr
ADI
ent SDS PAGE gels and detected for GFP protein expression. The control for exosome lo
ADI
ng was an anti-CETP antibody. XPack-GFP exosomes had very high amounts of GFP protein loaded as cargo.



Figure 1. XPack efficiently packages GFP into exosomes.

(Upper panel)
XPack-GFP localization during packaging.
(Lower panel)
Western blot showing robust lo
ADI
ng of GFP into the isolated exosomes.

Part 2: XPack-GFP Exosome Delivery to Target Cells

The XPack-GFP exosomes isolated as described above were then added to recipient HEK-293 cells to test for GFP delivery. Cells exposed to XPack-GFP exosomes were imaged by fluorescent microscopy, which showed subcellular GFP localization consistent with endocytosis of the XPack-GFP exosomes and delivery of the XPack-GFP protein cargo into the cell (Figure 2, left panels). Subsequently, these recipient cells were washed twice with PBS and cellular lysates were prepared for Western blot analysis using anti-GFP antibodies. Anti-GAPDH was used as a cellular protein lo
ADI
ng control. The Western blot data further verified the successful delivery of GFP proteins to recipient cells via the XPack exosome engineering technology (Figure 2, right panel).


Figure 2. XPack-GFP exosomes deliver GFP to target cells. (Left panel)
XPack-GFP localization during packaging.
(Right panel)
Western blot showing robust GFP delivery to target cells.