2 replies to this topic

[nirupakl]

hi all,

Our lab offlate has been having  this problem. We work on frog skin proteins and irrespective of the antibody used, we always see two band at size 29kDa region.One being very thick and the other thin. These two bands always crop up alongwith  the band that shows up in our region of interest.we are unable to solve this.The two bands are very prominent during Ponceau step too and perfectly show up irrespective of the protein  antibody-moniclonal or polyclonal.
can any one give some suggestions?

[merwyn]

Hai
    If that ptn is recombinant, then it may b due to premature termination.
    If the ptn is a native one (Probbly  directly from frog) the bands may b due to mature and preprossed ptn

[HomeBrew]

Are your primary antibodies conjugated, or do you use a secondary antibody to visualize them?  If you use a secondary Ab, perhaps it's cross-reacting with the protein(s) in the aberrant bands.