We require a minimum 20 pmol of blotted protein. 50 pmoles is very comfortable and generate unambiguous data.
|Protein size||µg for 20 pmoles|
We accept PVDF blots or purified lyophilised/solubilised proteins.
The protein should be electroblotted from 1D or 2D gels and stained with Coomassie Blue (R250). Following staining/destaining, blotted membranes should be rinsed thoroughly with deionized water. The sample should be as concentrated as possible (15 to 20 pmol / lane) and several gel lanes may be loaded and submitted. The whole membrane may be submitted with a copy showing the band(s) of interest or the bands may be cut out and submitted, please indicate band molecular weight.
30 to 150 µl of solution (H2O, Acetonitrile, Propanol). Pure lyophilised protein is also acceptable.
Protein should be free of detergent, salts and buffers. Please note that any amine-containing chemical present (e.g. Tris) will impair the adduction step.
Extra charge applies for this type of sample as we will run pre-purification at nominal fee to ensure complete removal of amine-containing impurities.
PVDF blots: well separated bands.
Liquid: Single purified proteins. Immunoprecipitation and affinity-column purified usually give good samples for sequencing.
In any case, a mandatory additional column purification step is applied to all incoming solution samples.