Fluorescence image based Protein-Protein Interaction analysis

15A Constitution Way, Woburn MA 01801,USA • T: 800.200.5459 F: 781.939.6963 • mblintl.com

International Corporation

Fluorescent Proteins

l ife. science.discovery.™

• Protein-Protein Interaction in living cells

• No need for optimizing linker!

• High-throughput drug discovery system

Fluoppi (Fluorescent based technology detecting Protein-Protein Interactions) is a novel technology used to detect PPI in living cells with a high signal to noise ratio. The result is a bright, clear analyzable image. Fluoppi detects PPI as the absence or presence of fluorescent foci during inhibition or induction, respectively. The Fluoppi advantage is the ease of the construction of the PPI detection system, there is no need for optimizing the linker.

inhibition

inductionPPI (-) PPI (+)

15A Constitution Way, Woburn MA 01801,USA • T: 800.200.5459 F: 781.939.6963 • mblintl.com

pAsh-MCL

Four types of plasmids

phAG-MCL

phAG-MNL

*hAG=humanized Azami Green

pAsh-MNL

Ash linker MCSCMV

AshlinkerMCSCMV

hAG linker MCSCMV

hAGlinkerMCSCMV

pUC ori

SV40polyA

Pcmv

Neomycin

f1 ori

SV40 ori

“Flexible”peptide linker (22 aa): N term.- NSADG GGGSG GSGGS GGGST QG ‒ C term.

phAG-MCLinker

Fluoppi employs an oligomeric assembly helper tag (Ash-tag) and

a tetrameric fluorescent protein tag (FP-tag) to form detectable

fluorescent foci when there are interactions between two tagged-

proteins of interest. Because each fusion protein has multiple Xs

or Ys, interaction between protein X and Y causes large lattice like

complexes where the fluorescence by X-FP is concentrated and

detectable as fluorescent foci.

Fluoppi tags are able to work in both N and C terminal fusion.

Plasmid vectors of Fluoppi consist of CMV promoter, Fluoppi

tags, flexible peptide linker, Multiple Cloning Site (MCS) and

Neomycin resistant gene. First, proteins X & Y of interest are

fused to FP-tag and Ash-tag respectively. Expression level of

tagged protein and foci formation efficiency depend on tag and

tag location Formation of foci is reversible so that they can be

dissociated and the fluorescent signal will spread over the cell by

PPI inhibitors, and vice versa by PPI inducers.

Versatile Protein Protein Interaction System Cytosolic protein-Cytosolic protein

Negative control PPI in cytosol

Nucleic protein-Nucleic protein

Negative control PPI in Nucleus

Cytosolic protein-Cytosolic protein

Negative control PPI at juxtamembrane

Code no. Product VolumeAM-8001 Fluoppi : Ash-hAG (Ash-MNL/MCL+hAG-MNL/MCL)* 10µg

AM-8201 Fluoppi : Ash-hAG [p53-MDM2] 10µg

AM-8202 Fluoppi : Ash-hAG [mTOR-FKBP12] 10µg

AM-8002M Fluoppi : Ash-Red (Ash-MNL/MCL + Monti-Red-MNL/MCL)

*This kit consists of four plasmids (pAsh-MCL, pAsh-MNL, phAG-MCL, phAG-MNL)

Coming soon!