Abgent's selection of bacterial and baculovirus
protein expression systems provides a choice for your application.
We will work with your specifications for quantity, bioactivity,
structure, post-translational modifications, and purity to express
proteins that meet your specialized requirements. Project status and data reports are monitored by our exclusive in-house custom service database tracking system.
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| Phase |
Time |
I. Cloning of a gene of interest into a bacterial expression vector1
- Amplification/isolation of the gene of interest out of a customer-supplied vector and subcloning it into a bacterial expression vector.
- Verification of the authenticity of the subcloned gene by restriction enzyme digest and sequencing.
- Maxi-prep of the recombinant vector DNA.
|
2 weeks |
II. Generation and identification of bacterial colonies expressing target proteins
- Transformation of recombinant constructs into high-efficiency expression bacterial strain.
- Mini-induction to over-express the target protein.
- Test for the expression of the recombinant protein by western blot (customer must provide appropriate antibodies) if desired.
|
1 week |
III. Large-Scale Culture
- One liter of bacterial culture will be induced with IPTG and harvested for protein purification.
|
2 days |
IV. Purification2
- Protein purification using Ni-NTA (for 6xHIS-tagged proteins) or glutathione (for GST-tagged proteins) beads at either native or denatured conditions.
|
1 week |
| Total |
4 weeks |
|
