Does anyone has experience of separating D2long and short form using SDS-Page? I have run the mouse brain tissue sample on SDS-page at 165 v for approximately 3 hrs to separate D2long and short form and blotted on PVDF membrane. Treated with rabbit polyclonal D2 antibody (Sc 9113) followed by secondary antibody. i could see two bands apporx. at 48 and another at 52 Kda. But my concern is that how to confirm whether those band are D2L and D2S? Any suggestion Please !!
I'm no expert in those proteins, but I'd say that confirmation of a band corresponding to a given protein would be achieved by combination of the molecular wheight determination (do those match the ones you wanted?), imunoblot (which seems to have worked), and in last resourse band cutting and sequencing/identification by mass spectrometry...
I just ran a very brief and not extensive search in uniprot and MWs of short and long D2 isoforms are aproximately 47 and 51kDa respectively, so it seems you've got them... :)
Rafael Felix Could you please tell me the procedure to check molecular weight of D2 isoform using uniprot...
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