Recombinant protein expression service

Transcription

1 309 EUROGENTEC 08 Catalogue > > 17 Recombinant protein expression service Customized protein expression 310 Sample and shipping requirements 312 Protein expression order form 313

2 Recombinant protein expression services 17 Customized protein expression service How do I start? Easy! Send us your gene sequence, vector sequence and required amount of protein to be expressed by to protein.expression@eurogentec.com You will receive a sequence analysis report by within the next 3 days. Is this service guaranteed? As all proteins are different and express at different levels, we cannot provide a guarantee that we will successfully express your protein. However you will only be invoiced for the services completed and you are free to exit the protocol at any step. Confidentiality Eurogentec maintains all projects in the strictest confidence. Should you require a material transfer agreement please us at info@eurogentec.com Then what happens? We will provide you with a free of charge report describing Expression in E. coli (prokaryotic) Purification of 1, 5, or 10 mg of your protein Choice of different protein tags Choice of purification levels, standard purity 80 % or high purity up to 90 % Customized service Competitive prices to fit your budget Whether you are interested in crystallography, NMR, protein-protein interactions, or activity assays, Eurogentec now also offers a protein expression service to advance your research needs. You choose your protein of interest, we will produce the protein for you, providing you with the purified material, the plasmid, and the expressing bacteria. Codon compatibility Vector compatibility Restriction enzyme analysis A strategy for the expression of your protein A price quotation for the proposed strategy An estimate for the time required to perform the services Protein expression Two options 1. We start from your mrna sample 2. We start from your cloned gene plasmid EUROGENTEC 08 Catalogue > > order@eurogentec.com Eurogentec s.a.s.u.

3 311 What do I receive? Report at the end of each step on - Codon usage - Vector compatibility - Housekeeping genes - Screening and sequencing - Small scale test expression - Large scale expression - Protein identity Purified protein to your requested scale and purity Technical support Your RNA or cdna sample Codon usage evaluation RT-PCR of your gene Ligation into pstaby or your vector Screen and sequence best clone Your cloned plasmid Custom protein expression services flowchart Delivery of your Protein and Plasmid Start Option I: protein expression from your mrna sample Purify protein and QC by SDS-PAGE and MALDI-TOF Large scale expression at 37 C or 28 C Test expression levels at 37 C, 28 C if necessary Transform into an E. coli production strain Start Option II: protein expression from your cloned plasmid Starting from your cloned plasmid we will Small scale protein expression evaluation - 10 ml at 37 C, rich-media - SDS-PAGE analysis to determine expression efficiency - Small scale expression results report Scale up to produce desired quantity of proteins - Protein expression scales from 1 to 10 mg - Purification against your desired tag - QC by MALDI-TOF and SDS-PAGE - Protein expression and QC results report Order references Protein expression Reference Description Days DG-PEXP-EVAL Codon usage, vector evaluation report 1 DG-PEXP-QCSM QC starting material 2 DG-PEXP-AMP Amplification 1 DG-PEXP-LITR Ligation, transformation and screening 10 DG-PEXP-SM37 Small-scale expression test, 37 C, rich media 8 DG-PEXP-SM28 Small-scale expression test, 28 C, rich media 10 DG-PEXP-LG37-01 Large scale protein expression, 37 C, 1-2 mg, 15 Starting from your mrna sample we will QC the material against housekeeping genes to ensure the integrity of the sample: - You receive a QC report PCR amplification and purification, includes primer design Ligation into the pstaby vector: - You receive a cloning report Screen for the correct clone and sequence the clone - You receive a screening and sequencing report Transformation of the recombinant clone into the E. coli production strain Continue with small scale protein expression evaluation below DG-PEXP-LG37-05 DG-PEXP-LG37-10 DG-PEXP-LG28-01 DG-PEXP-LG28-05 DG-PEXP-LG28-10 Large scale protein expression, 37 C, 5 mg, Large scale protein expression, 37 C, 10 mg, Large scale protein expression, 28 C, 1-2 mg, Large scale protein expression, 28 C, 5 mg, Large scale protein expression, 28 C, 10 mg, 15 EUROGENTEC 08 Catalogue > > order@eurogentec.com Eurogentec s.a.s.u.

4 Recombinant protein expression services 17 Sample and shipping requirements All samples must be accompanied by a document with the following information: investigator s name, address, telephone number, fax number and address, your institution s purchase order number. The order references for the services you have ordered. mrna sample requirements At least 5 µg of mrna are required for protein expression services, where the A260/A280 ratio is greater than or equal to 1.8. The concentration must be greater than or equal to 0.1 µg/µl and supplied in a nuclease-free packaging and buffer. Shipping your mrna preparation sample Ship your mrna sample on dry ice with an overnight courier. Please ship samples at the beginning of the work week to the address indicated below. Plasmid DNA for subcloning sample requirements Please resuspend 5 µg of highly purified vector DNA containing the gene of interest in sterile water or TE buffer to a final concentration greater than or equal to 0.1 µg/µl. Place in a microcentrifuge tube. Label the tube with the investigator s name, name of the recombinant plasmid, and the concentration of the DNA. Place the tube in a 50 ml conical tube labelled with the name of the recombinant plasmid, vector used, concentration of the DNA, and buffer used. Please indicate the name and restriction map of the clone and host plasmid, the restriction sites and strategy used for cloning, the size of the gene of interest and sequence data. Send at C. All samples Please complete the attached form. Shipping address Eurogentec SA Protein Expression Services LIEGE science park B-4102 Seraing Belgium Tel: How to order 1. Send an to protein.expression@eurogentec.com with your Option 1: gene sequence Option 2: gene sequence and vector sequence information Including the minimum quantity, minimum purity, and desired fusion tag to receive a project analysis and price quotation. 2. Fax your order form UK: FR: NL: BE and other countries: Eurogentec s.a.s.u.

5 313 Protein expression order form Your purchase order reference: VAT Nr. : EGT Quotation Number: If VAT exempt please include a copy of certificate INVOICE ADDRESS Institute/Company: Department/Lab.: DELIVERY ADDRESS (Please fill in delivery information if different from invoicing information) Institute/Company: Department/Lab.: Contact Person: Contact Person: Address: Address: Postal Code: Country: Phone: Fax: City: Postal Code: Country: Phone: Fax: City: Project name: Gene accession number: What is the starting material? q Genomic DNA q mrna q cdna already cloned What is the final use of the protein? q Antibody production q Crystallization studies q Functional studies q Other Minimum protein purity % Minimum protein quantity mg Protein tag requirements q 6xHis q GST q Other q Should not be included Have you already tried to express this protein? q No q Yes System used to express Are there any known difficulties associated with the expression of this protein? q No q Yes q Based on literature reports q Based on personal experience Additional Comments Eurogentec s.a.s.u.

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