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Custom Protein Services: Yeast Expression Systems
Insight Biotechnology provides a broad range of yeast protein expression services which can be tailored to your specific research needs. Choose any starting phase or combination. Insight Biotechnology takes a flexible approach towards custom protein services and information on this website is only a guide. Feel free to contact our team of protein expression experts with any further questions. Email, or telephone Insight Biotechnology on Tel: +44(0)208 3850303.
Please Click here for further information about other custom services offered by Insight Biotechnology.
Phase 1: Subcloning into Expression Vector |
Description |
Amplification/isolation of the gene of interest out of a customer-supplied vector and subcloning it into a suitable expression vector. |
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Verification of the subcloned gene by restriction digest and sequencing |
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Preparation of the recombinant vector DNA. |
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Required Materials |
5µg of plasmid containing gene of interest plus complete vector sequence in electronic format. |
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Deliverables |
1) 20µg of expression-ready recombinant plasmid containing the gene of interest |
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2) Final sequencing data |
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3) Detailed research report |
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Duration |
2 weeks; If insert > 2kb, one week additional per 1kb |
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Price |
£666; If insert > 2kb, £266 additional per 1kb; |
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Phase II: Generation and Identification of Expression Positive Clones |
Description |
Transformation of recombinant constructs into a suitable yeast strain. |
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Small scale induction to over-express target protein. |
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Test for recombinant protein by SDS-PAGE and western blot (customer must provide appropriate antibodies). |
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Required Materials |
5µg of expression-ready recombinant plasmid containing the gene of interest plus complete vector sequence in electronic format. |
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Deliverables |
1) Expression positive clones ( E. coli glycerol stock ) |
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2) SDS-PAGE and Western Blot data |
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3) Detailed research report |
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Duration |
2 weeks |
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Price |
£333 |
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Optional |
For His or GST tagged protein, you can choose to use our antibodies in stock, with an additional charge of £20. |
Phase III: Large-scale Culture |
Description |
One litre of yeast culture will be grown in induction medium suitable for the expression vector used. Recombinant protein expression is harvested for protein purification. |
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Test for recombinant protein by SDS-PAGE and western blot (customer must provide appropriate antibodies). |
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Required Materials |
Expression positive yeast clone together with a verified induction procedure |
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Deliverables |
1) Cell pellets from 1 litre fermentation |
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2) SDS-PAGE or Western Blot data |
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3) Detailed research report |
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Duration |
1 Week |
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Price |
£266 |
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Optional |
For His or GST tagged protein, you can choose to use our antibodies in stock, with an additional charge of £20. |
Phase IV: Protein Extraction & Purification |
Description |
Cell pellet harvested from 1 Litre of yeast culture after induction will be lysed with our optimised reagents and protocol. |
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Protein purification using appropriate affinity resin for 6xHis or GST-tagged recombinant proteins. |
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Purification can be performed either under native or denatured conditions (as desired by the customer). |
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SDS Page and Western Blot will be performed for quality testing. |
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Required Materials |
Cell pellet harvested from 1 litre of bacterial after induction |
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Deliverables |
1) Purified protein in standard or customer-specified solution |
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2) SDS-PAGE or Western Blot data |
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3) Detailed research report |
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Duration |
2-3 weeks |
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Price |
£999/L, enquire the price for larger volume |
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