Generon has teamed up with a core protein expression facility within the UK. We can offer you a complete protein expression service (with full customer and trouble-shooting support) from the accession number / sequence ID all the way through to biomolecular structural analyses using any of three high efficiency expression platforms: E. coli, insect and mammalian (cloning, small scale expression, scale-up, final purification, tag removal, customer-directed final analyses). From a biophysical analyses perspective, we can offer CD-spectroscopy, light scattering, mass spectrometry in addition to standard analysis tools. The service can be tailored to your specific requirements. Cloning Service:
Cloning (<1kb cDNA insert) from accession number- using either a cDNA library or an image clone as a template. We will design oligonucleotide primers to amplify the gene of interest and then clone this into the desired vector of choice. Constructs will be fully verified by DNA Sequencing. Prices for larger inserts available upon request. Buy online... Bacteria Phase I:
A small scale pilot study which includes: bacterial transformation, expression test using both IPTG and auto-induction methods, solubility testing, SDS-PAGE gel analyses and Western blotting. Buy online... Bacteria Phase II:
1 L scale up using the optimised conditions elucidated in Phase I to include protein expression, affinity purification, tag removal, second step purification (if required) and SDS-PAGE analyses of all protein fractions. Affinity tag will be removed if required by the appropriate enzyme digestion (eg. thrombin, TEV etc) and cleaved proteins purified by gel filtration or anion exchange chromatography. Buy online... Insect Phase I:
A small scale pilot study which includes: co-transfection of baculovirus DNA and transfer plasmid into Sf9 cells, P1 stock virus amplification, test infection in 6-well dishes using both Sf9 and Hi5 cell lines, at T48 and T72 hours with five concentrations of recombinant virus plus an infected control. SDS-PAGE analyses of cell fractions (total/soluble/pellet/secreted) and Western blot analysis. Buy online... Insect Phase II:
0.5-1L scale up using the optimised conditions elucidated in Phase I to include: amplification to P2 and P3 of virus stocks, infection of 0.5-1L of cells, protein affinity purification, tag removal, second step purification (if required) and SDS-PAGE analyses of all protein fractions. Affinity tag will be removed if required by the appropriate enzyme digestion (eg thrombin, TEV etc) and cleaved proteins purified by gel filtration or anion exchange chromatography. Buy online... Mammalian Phase I:
A small scale pilot study which includes: transfection into HEK293 EBNA cells and antibiotic selection of transfected cells. SDS-PAGE analyses of cell fractions (total/soluble/pellet/secreted) and Western blot analysis. Buy online... Mammalian Phase II:
0.5-1 L scale up using the optimised conditions elucidated in Phase I to Include growth of 0.5-1L culture, affinity purification, tag removal, second step purification (if required) and SDS-PAGE analyses of all protein fractions. Affinity tag will be removed if required by the appropriate enzyme digestion (eg. thrombin, TEV etc) and cleaved proteins purified by gel filtration or anion exchange chromatography. Buy online... Mammalian Biomolecular Analysis:
Analytical characterisation ensures the identity, purity, structural and conformational integrity of a protein. We perform a number of routine protein analyses throughout all project stages. We can also offer the following analyses for an additional charge: CD-spectroscopy, light scattering and mass spectrometry. Buy online...
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