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Protein Expression in Mammalian Cells

Even though many mammalian proteins have been successfully produced in bacteria, yeast, or insect cells, mammalian cell cultures are still the system of choice if mammalian-specific post-translation modifications are required. We conduct protein expression in mammalian cells via plasmid transient transfection, cell line establishment, retrovirus or lentivirus transduction, and recently introduced modified mRNA transfection. Also available are an array of cell lines that we have proven to be suitable for expression of different types of human or mouse proteins. We are able to use our fluorescent proteins and other expression tags for monitoring expression, or provide different drug-resistant genes if established cell lines are used for long term expression projects.


Phase I: Subcloning into Expression Vector

Phase II: (CMV Based): Expression in Target Cells

Phase III: (Retrovirus): Expression in Target Cells

Phase IV: Protein Extraction & Purification


Depending on your research needs, you may choose any starting phase or combinations.

Phase I: Subcloning into Expression Vector

Cat#  ABP-CS-PEMA001

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 Phoenix retroviral vector (provided by Allele Biotech, now including Orbigen).
  •  Verification of the subcloned gene by restriction digest and sequencing.
  •  Preparation of the recombinant vector DNA.

Required Materials: 5 ug of plasmid containing gene of interest plus complete vector sequence in electronic format.

Deliverables:

  1. 20 ug of expression-ready recombinant CMV based mammalian expression plasmid containing the gene of interest or 20 ug of Phoenix retroviral vector (ready for use) containing the gene of interest.
  2. Final sequencing data
  3. Detailed research report

Duration: 2 weeks; If insert > 2kb, an additional week per 1kb.

Price (USD): $999; If insert > 2kb, an additional $399 per 1kb.

Phase II: (CMV Based): Expression in Target Cells

Cat#  ABP-CS-PEMA002C

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: 20 ug 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 weeks

Price (USD): $1199

Optional: For His or GST tagged protein, you may choose to use our antibodies in stock for an additional charge of $29.

Phase III (Retrovirus): Expression in Target Cells

Cat#  ABP-CS-PEMA002R

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 ug of ready for use retrovirus expression plasmid containing the gene of interest plus the 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 (USD): $1499

Optional: For His or GST tagged protein, you may choose to use our antibodies in stock for an additional charge of $29.

Phase IV: Protein Extraction & Purification

Cat#  ABP-CS-PEMA003

Description: 

  • Cell pellet harvested after transient expression will be lysed with our optimized 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 (USD): $1500/L, discounts offered on larger volumes.