BioActive™ Fluorescence Protein Production Services

Seattle Genova is trying to offer our customers great biomarker protein production services that maintain the bioactivities of your proteins just as they were in nature. To this, we can offer both nature protein purification services and recombinant protein production services. We have formulate very specialized procedures to make sure the produced proteins will maintain their biological activities. And our well-developed bioassays can ensure extensive activity validations before we release the proteins.

Fluorescent proteins are units of a structurally homologous class of proteins that share the unique property of being self-sufficient to construct a visible wavelength chromophore from a sequence of 3 amino acids within their polypeptide sequence. It is common research practice for biologists to initiate a gene (or a gene chimaera) encoding an engineered fluorescent protein into living cells and then visualize the location and dynamics of the gene product utilizing fluorescence microscopy.

Nearly all endogenous proteins of interest can be fused to fluorescent proteins. Fluorescent proteins as genetically encoded equipment are used vastly by life scientists. The fluorescence-based assays have been applied to regulate a wide range of activities such as molecular dynamics and interactions, enzymatic activities, signal induction, cell health, and distribution of molecules, organelles, or cells.

Sample Preparation 

The target gene is cloned into expression vectors for the mammalian, insect or E.coli cells. The plasmid cDNA is extracted from the cells and transfected into 100 ml of E. coli or 50 ml of mammalian or insect suspension cells. 

Protein Purification 

The target protein is purified via affinity chromatography. The concentration and purity of the purified protein are determined using a bicinchoninic acid (BCA) assay and SDS-PAGE with Coomassie blue staining, respectively. The protein is desalted into 0.2 µm filtered 1x phosphate-buffered saline (PBS), pH 7.4. The protein can then be aliquoted, lyophilized, and labelled upon request.

Features of Our services 

√Expert Technical Support

Technical support is given by every scientist that develops the assays.

√Quality and User-friendly

Products are extensively assessed and validated before release so researchers need little-to-no time for assay optimization.

√High purity

With complete control over all quality testing, Seattle Genova can generate proteins to meet industry-leading purity specifications

√Low Endotoxin

Each new production lot of protein is assessed for endotoxin using the Limulus Amoebocyte Lysates (LAL) assay. Our standard endotoxin specification is an industry-leading <0.1 EU/ug.

Deliverables

√SDS-PAGE data 

√Western blotting data

√Bioactivity validation data

√Purified fluorescence protein

References

1.Smith, P.K.; et al. (1985). "Measurement of protein using bicinchoninic acid". Anal. Biochem. 150 (1): 76–85. DOI:10.1016/0003-2697(85)90442-7. PAID 3843705.

2.^ Olsen BJ, Markwell J (2007). "Assays for the Determination of Protein Concentration" (PDF). Current Protocols in Protein Science: 14–17.

3.Valeur, B.; Berberan-Santos, M. R. N. (2011). "A Brief History of Fluorescence and Phosphorescence before the Emergence of Quantum Theory". Journal of Chemical Education. 88 (6): 731–738.