Articles

Suspension Sf9 Cells from Monolayer Cultures

The transition to suspension Sf9 cells from monolayer cultures is not difficult but does depend on harvesting the attached cells at the right time. Too early and it is extremely hard to detach the cells from the culture flask. Too late and the cells will be in such poor condition that they may not recover when in suspension. Trying to describe in words how the monolayer culture should look prior

Product Launch – pOET8!

November marks the welcome of OET’s newest product – the pOET8 baculovirus transfer vectors (pOET8.VE1; pOET8.VE2; pOET8.VE3).  Compatible with any baculovirus expression system, including our own flashBACTM technology, the new range of pOET8 vectors are proven to help increase recombinant protein production in insect cells. The increased yield is achieved by allowing co-expression of foreign genes with a vankyrin expression cassette.  This gene cassette comprises the Campoletis sonorensis ichnovirus P-vank-1 protein

Reviving Cells from Liquid Nitrogen

The process of reviving cells from liquid nitrogen, or indeed from transport on dry ice, can be a little difficult on occasion.  Two events at OET recently have highlighted this. Firstly, we received a new cell line that came with a very complicated procedure for its culture – it is a mammalian species, quite different from your run of the mill HeLa’s and HEK293’s.  Its culture is known to be

Co-infection with Baculovirus Vectors for Multiple Protein Expression

A question that comes up from time to time is the use of co-infection with baculovirus vectors for multiple protein expression in insect cells.  The need arises if you have produced a number of baculovirus vectors that each synthesise a component of a multi-subunit complex, such as an enzyme or a virus like particle.  Intuitively, you know that it is probably necessary to add many virus particles per cell to

Baculovirus Transfer Vectors Compatible with flashBAC™

A question we are asked from time to time is to identify baculovirus transfer vectors compatible with our flashBAC™ system. We obviously nominate our own extensive range that is available but frequently someone has an old plasmid from years ago that they want to reuse. Originally they may have paired it with a linear DNA system such as BaculoGOLD or BacPAK6 or even wild type DNA where recombinant virus selection

Variable Expression of Multi-Component Protein Complexes

Variable expression of multi-component protein complexes is increasingly necessary as more sophisticated projects are attempted.  But how can this be achieved?  Most of our expression work is based on the baculovirus system, in particular our flashBAC™ vectors that permit easy recombinant virus generation in a one-step process.  These insect-specific viruses offer an ideal platform for producing a wide range of recombinant proteins including nuclear, cytoplasmic, membrane-bound and secreted examples. But

Transduction of whole porcine kidneys with BacMams

One of the unexpected offshoots of the baculovirus expression system has been its development as a vehicle for the delivery of genes to mammalian cells.  These vectors are otherwise known as BacMams.  Although baculoviruses are unable to replicate in mammalian cells, they can enter them via receptor-mediated endocytosis.  In Frederick Boyce’s original paper the Rous sarcoma virus promoter was used to express beta-galactosidase in the HepG2 human liver cell line. 

Expression Systems Media

Expression Systems insect cell culture media OET is pleased to offer a wide range of Expression Systems Insect cell culture media, from the very popular ESF 921, to specialized insect transfection media. Expression Systems’ media are easy to adapt your cells into and the insect specific media gives great protein yield, when compared to rival media, (comparative data to follow soon). ESF921 insect culture media OET use ESF 921 for

High titre virus stocks

Following our high titre virus stock blog, here are some OET products that will help you get the best results: baculoQUANT all-in-one is our QPCR kit that enables accurate and easy virus titring ESF 921 insect cell culture medium is specially designed for insect cells, improving the condition and performance of your cells and thus increasing virus production Super Sf9 insect cells are an optimised cell line, increasing virus production

Cells at Christmas

So, you have had the Christmas Party and hopefully are looking forward to the holiday season.  Many labs shut down between Christmas and New Year and you may not return until January 4th.  In all the build up to the festive season have you remembered everything?  Aunt Hilda’s bottle of lavender water and Uncle Horace’s box of cigars may be already wrapped and under the tree but aren’t you forgetting

Cell Culture Passage History and Stress

No, not how you feel when the cells don’t behave! This relates to keeping your cell cultures in a happy state. Cell passage history Each time you sub culture your cells you probably faithfully record on the new flask various bits of information such as the name of the cell line, date, the medium used and passage number.  This last item ideally relates to the number of times the cells

Cell Culturing OETips, information and manuals

Insect Cell Culture Manual download For Sf9 and Sf21 cell revival guide, please click here For Sf9 cell information, please click here For specialised plaque assay Sf21 information, please click here For Super Sf9 information, please click here How are they growing? Follow our OETips guide to cell culturing here.

How can protein expression be made easier? flashBAC™

The flashBAC™ system flashBAC™ is a revolutionary platform technology for the production of recombinant baculoviruses in insect cells. It comprises a parental AcMNPV genome that lacks part of an essential gene (ORF 1629), which removes the need to separate recombinant virus from parental virus. We offer several different types of flashBAC™, each with different set of deletions to enhance the yield of recombinant proteins. Yes, also those difficult ones that

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