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Transcellular transport

From Molecular Biology Wiki

Antibody Delivery System

Contents [hide]

   * 1 Author
   * 2 Background
   * 3 Protocol
   * 4 Acknowledgements

[edit] Author Ivan K - Cell Biology & Immunology Laboratory

[edit] Background Antibodies are not able to pass through plasmic membranes of live cells towards potential intracellular targets. Thousands of antibodies have been developped as tools for the scientists. Their intracellular delivery opens new fields of investigation for both research and therapeutic applications.

[edit] Protocol Antibody delivery protocol is provided for a 14-well plate 1- Add 2 µL of ImmunoCellin at the bottom of a microtube 2- In an other microtube, dilute 1 µg of your BSA free antibody in 10 µL of PBS 1X solution 3- Transfert the diluted antibody solution into the ImmunoCellin containing microtube and mix by pipetting 4- Incubate 15 minutes at room temperature 5- Add 90 µL of serum-free culture medium into the microtube and transfert the 100 µL mixture drop-wise onto the cell 6- Incubate at 37°C in a CO2 incubator 7- Analyse antibody delivery 5h-48h hours later depending on your experiment type For fluorescent microscopy, wash cells with PBS and fix them with 2-4% paraformaldehyde or formalin.

[edit] Acknowledgements BCC's R&D and technical support

Passive Active Transport

Use ATP as a source of energy. Na / K ATPase

Transcellular transport
 

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