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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

Procedure ?
Bring the proteins to the Platform with the form to complete.
 
The screening is made on the Synquad robot Cartesian (sitting drop method only) or on the Mosquito robot (both sitting or hanging drop). The Cartesian robot produces only 200 nl drops for the screening. The Mosquito robots proposes drops from 100nl to 1000 nl. The plates are sealed and preserved at 18°C in the Formulatrix Robot (with an imaging scheduling) until obtaining crystals. Once the crystals obtained and after checking that they are protein crystals well (see diffraction of the X-rays), the conditions of crystallization can be refined with the possible assistance of the Matrixmaker robot (see the Matrixmaker service).

 Kits available to the Platform
 
The initial screening is 6 plates of 96 different conditions : 576 different solutions mixture:
 
Kit Crystal screen1 and  2 –Hampton – HR2-110 and HR2-112
 
Kit Wizard 1 and 2 –Jena Biosciences  – CS-315
 
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 PEGion1 et 2  – Hampton:
PEGion 1 Screen – HR2-126
PEGion 2 Screen – HR2-098
 
Nextal kits – Qiagen :
JCSG Core 1   – 130924
JCSG Core 2   – 130925
JCSG Core 3   – 130926
JCSG Core 4   – 130927
 
Some other more specific  kits at your disposal :
 
Additives Hampton – Hampton  – HR2-428
 
Kit Index – Hampton  – HR2-144
 
Kit Proplex – Molecular dimensions    – MD1-38
 
Kit Natrix – Hampton  – HR2-116
 
Kit Memstart/Memsys – Molecular dimensions  – MD1-04ROWORLD
 
Kit JBScreen Membrane 1,2,3 – Jena Biosciences :
JBScreen membrane 1   – CS-301L
JBScreen membrane 2   – CS-302L
JBScreen membrane 3   – CS-303L
 
Kit JBScreen Cryo 01 – Jena Biosciences  – CC-103
 
Kit MembFac – Hampton  – HR2-114
 
Kit memgold – Molecular dimensions  – MD1-39

Access Protocol

Procedure of request for crystallization : 

1- Download the form  on the WEB page (PDF file), complete it and bring it io 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.

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 frehly as possible.

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

Academic research Cristallization Service :

⇒    CARTESIAN robot : 15€ per plaque and tested sample.

⇒    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 Pasteur Institute Platfom and the company for crystallization facilities. Please contact Ahmed Haouz (pf6@pasteur.fr)