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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.

 

Verification of the subcloned gene by restriction digest and sequencing.

 

Preparation of the recombinant vector DNA.

 

 

Required Materials

5µg of plasmid containing gene of interest plus complete vector sequence in electronic format.

 

 

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.

 

2) Final sequencing data

 

3) Detailed research report

 

 

Duration

2 weeks; If insert > 2kb, one week additional per 1kb

 

 

Price

Enquire


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.

 

Harvest and lysis of cells (usually after 2-3 days).

 

Test for recombinant protein by western blot (customer must provide appropriate antibodies), if desired.

 

 

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.

 

 

Deliverables

1) Cell pellet from 107 cells after transient expression.

 

2) Western Blot data (if desired)

 

3) Detailed research report

 

 

Duration

1 week

 

 

Price

Enquire

 

 

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.

 

Retrovirus isolation and titration.

 

Infection of target cells for transient recombinant protein expression (107 cells).

 

Harvest and lysis of cells (usually after 2-3 days).

 

Test for recombinant protein by SDS-PAGE and Western blot (customer must provide appropriate antibodies), if desired.

 

Excess of retroviral stock may be provided to customer.

 

 

Required Materials

20 µg of ready for use retrovirus expression plasmid containing the gene of interest plus complete vector sequence in electronic format.

 

 

Deliverables

1) Cell pellets from 1 liter fermentation

 

2) SDS-PAGE or Western Blot data

 

3) Detailed research report

 

 

Duration

1 week

 

 

Price

Enquire

 

 

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.

 

Protein purification using appropriate affinity resin for 6xHis or GST-tagged recombinant proteins.

 

Purification can be performed either under native or denatured conditions (as desired by the customer).

 

SDS Page and Western blotting will be performed for testing protein expression.

 

 

Required Materials

Cell pellet harvested after transient expression

 

 

Deliverables

1) Protein solution in standard or customer-specified buffer.

 

2) SDS-PAGE or Western Blot data

 

3) Detailed research report

 

 

Duration

2-3 weeks

 

 

Price

Enquire

 

 

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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