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Mammalian Expression Systems
Insight Biotechnology provides a broad range of protein expression services using mammalian systems, 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.
Available services are as follows.
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 CMV promoter based high-level expression vector or a retroviral 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 CMV based mammalian expression plasmid containing the gene of interest or 20µg of ready for use retroviral vector 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; |
Phase II: (CMV Based): Expression in Target Cells |
Description |
Transfection of recombinant vector into target cells (107 cells. Choose from 293, CHO, COS-7, and NIH 3T3) for transient expression. |
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Harvest and lysis of cells (usually after 2-3 days). |
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Test for recombinant protein by western blot (customer must provide appropriate antibodies), if desired. |
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Required Materials |
20µg of ready for use CMV based recombinant mammalian expression plasmid containing the gene of interest plus complete vector sequence in electronic format. |
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Deliverables |
1) Cell pellet from 107 cells after transient expression. |
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2) Western Blot data (if desired) |
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3) Detailed research report |
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Duration |
1 week |
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Price |
£800 |
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Optional |
For His or GST tagged protein, you can choose to use our antibodies in stock, with and additional charge of £20. |
Phase II (Retrovirus): Expression in Target Cells |
Description |
Transfection of recombinant vector into amphotropic or ecotropic packaging cells (107 cells) for transient expression. |
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Retrovirus isolation and titration. |
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Infection of target cells for transient recombinant protein expression (107 cells). |
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Harvest and lysis of cells (usually after 2-3 days). |
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Test for recombinant protein by SDS-PAGE and Western blot (customer must provide appropriate antibodies), if desired. |
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Excess of retroviral stock may be provided to customer. |
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Required Materials |
20 µg of ready for use retrovirus expression plasmid containing the gene of interest plus complete vector sequence in electronic format. |
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Deliverables |
1) Cell pellets from 1 liter 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 |
£999 |
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Optional: |
For His or GST tagged protein, you can choose to use our antibodies in stock, with and additional charge of £20. |
Phase III: Protein Extraction & Purification |
Description |
Cell pellet harvested after transient expression 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 blotting will be performed for testing protein expression. |
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Required Materials |
Cell pellet harvested after transient expression |
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Deliverables |
1) Protein solution in standard or customer-specified buffer. |
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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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Optional: |
For His or GST tagged protein, you can choose to use our antibodies in stock, with and additional charge of £20. |
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