= Western blot antibody specificity control
A known target-containing sample is a positive comparison for <Western blot antibody detection>. A matched sample with <gene deletion> supplies a negative comparison: disappearance of the proposed band supports the assignment, while a persisting band indicates binding unrelated to the deleted target. Deletion must actually remove the recognized <protein>; changes in other <proteins> can complicate the comparison. Omitting the <primary antibody> checks binding or background from the <secondary antibody> and detection reagents, but cannot exclude binding of the <primary antibody> to an unintended <protein>. Independent <antibodies> recognizing different <epitopes> add evidence. <Considerations when quantitating protein abundance by immunoblot by Alicia A. McDonough and others>[McDonough and others, “Consider the Specificity of the Antibody for the Target”, Figure 2: positive tissue, knockout tissue and antibodies to different termini]{ref}
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