Effective services and products dedicated to various research programs targeting the microtubule cytoskeleton (oncology, etc.)

Introduction

Microtubule cytoskeleton plays several major roles in cells. It is particularly important during mitosis, when microtubule-based bipolar spindle assures chromosome partitioning between daughter cells. Failure to properly segregate chromosomes in mitosis is frequently associated with cancerogenesis and tumour progression.

Due to its importance during mitosis, the microtubule cytoskeleton turned out to be a natural target for many of the currently available anti-proliferative drugs. Most of the currently known anti-mitotic drugs act by inhibiting tubulin assembly or depolymerisation, arresting in mitosis and eventually killing fast dividing (cancer) cells.

Services Offerings

A/ Products available

• Highly purified brain tubulin (purity >99%),
• Fluorophore-labelled bovine brain tubulin (Rhodamine, FITC, Cy3 and Alexa^TM fluorophores),
• Other fluorophores may be coupled to tubulin on request.

Two major reasons to test your compound in a tubulin polymerization assay:

a) You have identified an antiproliferative molecule and wish to make sure it affects tubulin

b) You have a hit/lead molecule-candidate drug and wish to exclude that your compound interferes with tubulin

--> Key Benefits

• While the first case (a) is mostly related to cancer drug research, the second (b) situation is important to all drug development projects. Indeed, a quick and reliable tubulin polymerization assays, carried out with the hit and/or lead molecule, can potentially save a lot of time and money.
  
• Interference with tubulin and/or microtubules can be a killer problem for any potential drug candidate.  It is thus imperative to identify and eliminate tubulin interfering compounds at the earliest stage of a drug development program.
• Tubulin screening should not be overlooked, taking into account that in some commercial small molecule libraries the percentage of chemicals that interfere with tubulin and or microtubules could be as high as 5%.

Cell-based assay for small molecules interfering with microtubule cytoskeleton

Indirect immunofluorescence on fixed HeLa cells. Microtubules are in green, nuclei in red. Control (DMSO-treated) cells are on the left, drug-treated cells with depolymerized microtubules are on the right.

This assay allows to estimate:

a)  effects of test compounds on cell proliferation
b) mitotic index
c) organization of the microtubule cytoskeleton in different stages of the cell cycle

--> Key Benefits

Cell-based microscopy assay, which allows evaluating the morphology of the microtubule cytoskeleton in drug-treated cells (interfering with other than tubulin proteins and these effects can be overlooked using turbidimetry).

Custom production of monoclonal antibodies (mABs) for microscopy

Services available:

a) developing microscopy-friendly mABs against your favourite protein (YFP) (tests of supernatants from antigen-specific hybridomas on fixed cells)
b) isolation of hybridomas with predicted intracellular reactivity
c) un-covering unique populations of the same protein (conformational epitopes).

--> Key Benefits

• Indirect immunofluorescence microscopy (IIM) is a powerful technique avoiding you the usual following problem:
  
  • Antibodies developed using reactivity in ELISA and immunoblotting, often fail to recognize their target(s) in fixed cells or give a staining incompatible with localization(s) observed in live cells (for example expressing a fluorescent fusion of a protein).

• In microscopy, mABs have some additional advantages aver polyclonal reagents:
  
  • mABs are usually more specific;
  • mABs give less background;
  • mABs produce more reproducible results.

Applications

• Cell biology
• Developmental Biology
• Immunochemistry and immunocytochemistry

Commercial Opportunities

Floralis is currently looking for industrial partners who may benefit from this exciting new technology as part of their R&D programs.

Contact
Florence Alesandrini
+33 (0)4 76 00 78 35
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