SERCA2a: various bands

[sammieC]

Hiya! I was wondering if anyone has used the anti-serca antibody on rat cardiac homogenates, and how your blots look like when they came out from the developer?
I am using Anti-Serca antibody from Millipore, and I have 'boiled' my samples for 5minutes...Used 9% gel..
Does anyone just get one band at ~ 110KDa? or, like me- get bands at ~110KDa and other bands elsewhere on the blots too? This is confusing me as I am not sure what these other bands are?
Please help me! thanks so much in advance..

[Jesi]

Hi Sammie! I usually use a-SERCA antibody on rat cardiac homogenates for western blott. When I boiled them for 5´, the band on 110kDa dissapear (I have no reasson for that), so I kept them on ice until I put them inside the gel. Even this, same inespecifics bands still appear. I have another problem, maybe you can help. Same samples appear o disappear depending on the experiments although they were prepair in the same conditions.