2 replies to this topic

[kiwidiwi]

Hi all, I've been struggling with actin lately. The actin bands - repeatedly, with different sample (within the same screen) and different runs/transfers,
the actin gives me weird results. (attached pic). So far people speculate it's a transfer bubble problem but all the other proteins on the same blot are near perfect whereas only actin is disastrous with distinctively shaped band. I tried fresh antibodies but no improvements.

I am suspecting my running/transfer buffer has gone bad since I reuse them, (if this is the case, why it would just mess up actin is a mystery to me)
and the next thing I could think of is stripping since I strip (commercial buffer) for 20 minutes and then reblot (4C, o/n) for actin.

Has anyone ever had a similar problem? or other suggestions?

Attached Thumbnails

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[phage434]

Is this an SDS gel or native gel? Actin forms strong multimers, which you could be seeing in a native gel.

[kiwidiwi]

Unfortunately it's SDS gel. I have no idea why this is just happening to actin !