For Laboratory Research Use Only. Not for Use in Clinical Diagnosis or Therapeutic Procedures.

Antibody delivery reagent

 Product Name  Cat # Size Price
GenomONE-CAb EX    AB001EX 1kit (HVJ-E 1 vial set) distributor website
AB004EX 1kit (HVJ-E 4 vials set) distributor website

  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 de
livery 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 stimulationAntibody ()                   

                   
                                                    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 2-8C.
 
n [Reconstituted HVJ-E (HVJ-E suspension)] :  Requires refrigerated storage (2-8C) 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 (2-8C) for up to 2 weeks.
                                                               
        For extended storage, freeze in working aliquots at -80C 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)

 

 

 

 

 

 

 

 

Copyright  (C)  2010-2013  ISHIHARA SANGYO KAISHA, LTD. All Rights Reserved.