I am trying to Generate polyclonal antibodies towards the amyloid-like species of glucosylcerebrocide (GlcCer). Rabbit antisera were collected and used against this GlcCer. I have prepared GlcCer amyloid sample in PBS. Now I am trying to find out the binding affinity of this rabbit antibody with GlcCer sample through DOT Blot assay. My experimental design is as follows:

Control lane: PBS+ rabbit antisera

Sample: PBS+GlcCer+rabbit Antisera

I found the same signal from both lanes. How do I optimize this error?

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