- Further Characterisation
- Transfectant Preparation
- Western Blotting
- Staining Projects
Transfectant Preparation
We always recommend that any ELISA positive supernatants are further characterised by testing against mammalian cells that have been induced to express the target protein by transfection with the corresponding gene, if appropriate. Other screening methods can also be employed This helps to ensure the specificity of any antibodies raised. OxFabs can produce an expression construct and perform the transfections or we can use reagents supplied by the customer. The schedule below gives an idea of a minimum time scale for the production of transfectants from gene name only.
Appropriate IMAGE clone identified, ordered and sequence verified
8 days
PCR primer design and amplification of antigen from IMAGE clone
4 days
Antigen cloned into mammalian expression vector
9 days
Mammalian cells cultured to optimal density for transfection
4 days
Transfections optimised for expression construct of interest
2 days
Cytocentrifuge preparations of transfected cells produced and stored until required for screening
1 day
Transfected cells prepared for antibody screening can be produced within one month. If the customer has requested recombinant protein to be made by us the IMAGE clone will already have been ordered and sequenced by us reducing the time required to produce transfected cells.