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© Ahmed Haouz
Cristaux d'une protéine de Mycobacterium tuberculosis produits dans le cadre du Grand Programme Horizontal sur la Tuberculose à l'Institut Pasteur. La caractérisation structurale de protéines mycobactériennes aide à une meilleure compréhension de la physiologie et de la pathogénicité des mycobactéries et fournit un point de départ pour la conception de nouveaux agents antibactériens.
Scientific Fields
Diseases
Organisms
Applications
Technique
Starting Date
12
Mar 2015
Status
Ongoing
Members
3
Structures
1

About

Application process of crystallization at 18°C or 4°C

In the platform we can setup high-throughput crystallization experiments (18°C or 4°C) with the vapour diffusion (sitting or hanging drop), the microbatch, and the cubic phase methods. The screening can be made with the Mosquito  or with the Douglas ORYX-8 robots. These robots can dispense drops from 100nl to 2µl. The crystallization plates will then be stored at 18°C or 4°C in a Formulatrix® Robots (an automated imaging system for protein crystallization). The crystallization drops will automatically be imaged every day. If successful results are obtained, the crystals will be reproduced and optimized using crystallization solutions generated by a Matrix Maker automated formulation system (Emerald BioSystems®) (see the Matrixmaker service).

Bring the proteins to the Platform with the filled form

 Kits available on the Platform
The initial screening is 6 or 7 plates of 96 different conditions among the following kits:
Kit Crystal screen 1 and  2 – Hampton: HR2-110 and HR2-112
Kit Wizard 1 and 2 – Jena Biosciences:  CS-311 and CS-312
Kit MDL 1 and 2 – Molecular dimensions:
Kit Structure screen 1   – MD1-01
Kit Structure screen 2   – MD1-02
Kit JBS1-4 –Jena Biosciences:
JBS Screen 1 bulk    – CS-101L
JBS Screen 2 bulk    – CS-102L
JBS Screen 3 bulk    – CS-103L
JBS Screen 4 bulk    – CS-104L
Kit JBS 5-8 – Jena Biosciences:
JBS Screen 5 bulk   – CS-105L
JBS Screen 6 bulk   – CS-106L
JBS Screen 7 bulk   – CS-107L
JBS Screen 8 bulk   – CS-108L
Kit Salt RX HT – Hampton:   HR2-108
Kit PEGion 1 and 2  – Hampton:
PEGion 1 Screen – HR2-126
PEGion 2 Screen – HR2-098
Crystal Screen Cryo 1 and 2:   HR2-122 and HR2-121
Kit Index – Hampton:  HR2-144
Kit Proplex – Molecular dimensions:  MD1-38

Nextal Classic suite – 1054292

 Some other more specific  kits at your disposal :
Kit Memstart/Memsys – Molecular dimensions:  MD1-04ROWORLD:
Memstart – MD1-21
Memsys – MD1-25
Kit JBScreen Membrane 1,2,3, 4 – Jena Biosciences:
JBScreen membrane 1   – CS-301L
JBScreen membrane 2   – CS-302L
JBScreen membrane 3   – CS-303L
JBScreen membrane 4   – CS-304L
Kit MembFac – Hampton:   HR2-114
Kit Natrix – Hampton:  HR2-116
Kit memgold 1 – Molecular dimensions:  MD1-39
Kit memgold 2 – Molecular dimensions:  MD1-63
Additives Hampton – Hampton:  HR2-428
Additives MEMadvantage:  MD1-70
And some homemade kits:

Access Protocol

Procedure of request for crystallization : 

1- Download the form  on the WEB page (PDF file), complete it and bring it to the Platform with the proteins.

2- Contact the platform  to specify the technical details in particular the buffer of conditioning and concentration of protein to be crystallized (the sample concentration must not be less than 5mg/ml). We need 40 µl of sample to set up 200 nl drop for each crystallization plate.

3- Provide the samples according to the planning. The date has to be booked BEFORE beginning a purification in order to screen the protein as freshly as possible.

The costs of the consumable are in charge of the laboratories.

Academic research Cristallization Service :

⇒    MOSQUITO robot (sitting drops): 20€ per plaque and tested sample.

⇒    MOSQUITO robot (hanging drops): 33€ per plaque and tested sample.

Private Companies : A contract has to be established between Institut Pasteur and the company. Please contact Ahmed Haouz (pf6@pasteur.fr)