|
GenomONE-CAb EX is a tool for efficient introduction
of IgG antibodies into
living cells using HVJ-Envelope (HVJ-E) vector for
the purpose of analysis
of the functions of cells and intracellular
proteins.
What's
HVJ Envelope (HVJ-E)?
[Features]
Because antibodies cannot enter cells, past experiments using
antibodies focused primarily on extracellular
molecules.
If
target molecules in living cells can be exposed to antibodies, it will be
possible to pursue new
dimensions of research related to cell function analysis, exploration
of target molecules for disease diagnosis
and
treatment, and so on.
GenomONE-CAb
EX
Antibody Delivery Reagent is a
next-generation tool for antibody
introduction into cells which can meet following
needs.
n
Analysis of intracellular function
n
Screening of antibodies reacting to intracellular antigens
n
Advantages of intracellular antibody introduction (differences from existing knockout method)
[Principle]
With this kit, antibody can be incorporated into the
HVJ Envelope (HVJ-E), a transfection tool making use of
the membrane
fusing ability of inactivated Sendai virus (HVJ: Hemagglutinating Virus of
Japan). If cells are
treated with HVJ-E including antibody, it will be
possible to achieve efficient introduction of IgG antibodies into
the
cytoplasm. This kit provides a totally new methodology for experiments,
overcoming the difficulties involved in
experiments using conventional
lipid-based reagents by which antibodies are introduced into cells by
means of
endocytosis.
With this system, the IgG antibody
incorporation enhancer, which was developed by our company,
improves
the efficiency of inclusion
of IgG molecules into HVJ-E markedly compared to the existing
HVJ-E
vectors
(GenomONE-Neo
EX ). Thanks to this feature, this
system allows efficient introduction of
IgG molecules into the cytoplasm.

Papers related to
GenomONE-CAb
EX
published by our
company.
Kondo
Y. et al.: Efficient delivery
of antibody into living cells using a novel HVJ envelope vector
system.
J. Immunol. Methods, 332,10-17(2008).
[Applications]
Examples of antibody introduction into
living
cells
|
Intracellular
delivery of anti-NPC
monoclonal antibody

HeLa S3
Hs68
Raw264.7
(ATCC-CCL 2.2)
(ATCC-CRL1635) (ATCC-TIB-71)
Actin filaments were stained
stained with phalloidin (red)
The
anti-NPC (nuclear pore complex
protein)
antibody, introduced into living
cells, moves onto
the nuclear membrane, holding its activity, where it binds specifically
to the antigen (resulting in
a ring-shaped
chromatic response of the nuclear membrane)
F (ab’)2
fragment
should be used as the fluorescence-labeled secondary antibody.
If whole antibody is used,
non-specific
binding may take place, making it impossible for specific chromatic
responses to appear.
|
|
Inhibition of
nuclear translocation of NF-κB

Non-stimulation/
Antibody(-)
LPS stimulation/Antibody
(+)

Control
antibody Anti-NF-κB antibody
Anti-NPC antibody
Introduction
of anti-NF-κB antibody or anti-NPC antibody to (red
arrows) Raw 264.7 cells
inhibited nuclear translocation of NF-κB by LPS
stimulation (yellow arrows).
(observed by
confocal laser scanning microscopy).
Inhibition of Nitric Oxide production
Introduction of anti-NF-kB antibodies or anti-NPC antibodies to
Raw 264.7 cells led to significant
inhibition of nitric oxide (NO) production.
(This experiment was performed as joint research with
Immuno-Biological Laboratories Co., Ltd.
We express our heartfelt thanks
to Dr. Kiyoshi Ishikawa and the other individuals involved.)
|
[Specification]
|
Cat. #
|
HVJ-E
(inactivated
HVJ)
Freeze-dried
0.26
mL /vial
(when reconstituted)
|
Reagent I
(antibody
incorporation enhancer)
Freeze-dried
0.26
mL/vial
(when reconstituted)
|
Reagent II (incorporation
reagent)
0.3
mL/vial
|
Reagent
III
(introduction
enhancer)
1
mL/vial
|
Buffer
(for suspension and
dilution)
6.5
mL/vial
|
|
AB001EX
|
1 vial
|
1
vial
|
1
vial
|
1
vial
|
1
vial
|
|
AB004EX
|
4 vials
|
4
vials
|
1
vial
|
4
vials
|
1
vial
|
[Storage]
n [Freeze-dried
HVJ-E and Reagent I]
:Moisture can reduce the activities.
Refrigerated at 2-8°C,
sealed in an aluminum package.
Keep dry.
n [Reagent
II,
III
and Buffer] : Refrigerated at 2-8°C.
n [Reconstituted
HVJ-E (HVJ-E
suspension)] : Requires refrigerated storage (2-8°C)
and should be used
within 2
weeks. Since thawing of frozen suspension can reduce
activity,
the suspension should not be stored frozen.
n [Reconstituted
Reagent I
(Reagent
I solution)]:
For continuous use, store in a refrigerator (2-8°C)
for up to 2 weeks.
For extended storage,
freeze in working aliquots at -80°C
for
up to
3 months. Thawing after freezing is possible
only once.
[Role
of each reagent]
n [Freeze-dried
HVJ-E]: The main frame of the vector into which the molecule to be
transferred is included.
It fuses with the cell membrane, allowing the
target molecule to be introduced into
the cytoplasm.
n [Reagent
I (antibody incorporation enhancer)]: A protein with high binding affinity
with HVJ-E and IgG
antibody
which increases incorporation efficiency of
antibody molecule into HVJ-E.
n [Reagent
II
(incorporation
reagent)]: Increases permeability across the HVJ-E membrane and
facilitate
incorporation of antibody.
n [Reagent
III (introduction
enhancer)]:
A positively-charged peptide which increases affinity between
the
IgG-bearing HVJ-E (HVJ-E vector) and the cell (or tissue)
and thus increases the efficiency
of delivery.
n [Buffer]:
Neutral buffer of physiological concentration used for suspending or
diluting HVJ-E or other purposes.
[Frequency
of use]
If
used with the method described in this package insert, the product can be
used for IgG antibody delivery as follows.
With
a 24-well plate
AB-001EX (HVJ-E:1 vial): 25
reactions
AB-004EX
(HVJ-E:4 vials): 100
reactions
With
an 8-well chambered coverglass for fluorescence observation
AB-001EX
(HVJ-E:1 vial):
50
reactions
AB-004EX
(HVJ-E:4 vials):
200
reactions
[Related
Technical Information]
[Related
product]
|
Transfection
kit
(For
introduction of DNA, siRNA and protein)
|
GenomONE-Neo
EX |
|
Cell fusion kit
[For preparation of hybridomas
(monoclonal antibodies) and research on development,
differentiation, and breeding (transplantation and replacement
of nuclei)]
|
GenomONE-CF
EX |
|
|
Order
(distributor) |
|
Contact us |
| Reference
article |
Related Technical
Informations

Flyer

Instruction manual
Safety Information

Application data sheet (No.1)

Application
data sheet No.2

Application data sheet No.3

Reference list
(in vitro)
|
|