Discriminating heterogenity

Hi all,

I’m working on a heterogenous sample, which I is a mixture of the target protein with one or two antibodies.

I’ve tried some methods (Hetero refinement, 3D classification) to fully separate the sample either with one antibody or two antibodies.
But when I check the map that I think it has one antibody at low threshold, there is still some signal of another antibody in the map…

Is there a way to completely separate the two?

Thank you for checking out.

nope, keep going. providing GOOD references for completely 1 vs 2 antibodies bound when doing het refinement should work after a few iterative rounds (take the ones classified as singly-bound, redo het refine with singly/doubly bound references, etc.). if there is a prevalent discriminating view in 2D this is possible also. so far there is no complete, correct, final separation outcome in this field. the tools are there but require user experience and persistence.

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Thank you for the idea. I’ll keep on pushing as your suggestion!