I intended to check for my target protein in the protein extract from maize leaves. I used anti-myc-AP Antibody (Invitrogen) to detect the interested protein which had cMyc peptide. After developing I was expecting only one band that is my target protein, but multiple bands were observed.
Does anyone have any suggestion regarding this problem?
Since, it's an extract, multiple bands may be due to non-specific binding. Try increasing the dilution of the antibodies.
Your protein may appear in several forms, either as alternative splicing, modifications, or you might have proteins with similar enough epitopes. since you didn't post the image of the gel, it's difficult to suggest what could be possible.
Thank you Himanshu and David, you can found the gel image in the attachment file. The arrow indicated my target protein.
Decreasing the antibodies concentration seems a good idea.
If you are getting non specific multiple bands with higher exposure time you can try lowering your exposure time while getting the signal . May be the antibody is not specific to your protein of interest cz sometimes I also experienced multiple bands .
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