b'Choosing a secondary antibodySelecting an appropriate secondary antibody is a critical step in the western blot detection procedure. A careful assessment of the needs of the experiment is required. The stepwise considerations below are useful in the process of choosing a secondary antibody.1 Determine origin of the primary antibody. 2 Select an appropriate host species andExample 1mouse monoclonal IgG1 target reactivity for the secondary antibody: Potential secondary antibody for example 1 Example 2rabbit recombinant IgG goat antimouse IgG1 Potential secondary antibody for example 2goat antirabbit IgG3 Consider cross-reactivity or specificity4 Detection method.issues of the secondary antibody.Labelappropriately conjugated to theCross-adsorbedfor multiple-labelingcorrect enzyme, tag, or fluorophore for applications or when using samples withthe chosen detection methodendogenous antibodies Specificitybinds to unique fragments, classes, or chains of the primary antibody 5 Consider requirementsof the6 Visit thermofisher.com/secondaries supplied secondary. for a complete list. Supplied statesuch as sterile liquid or lyophilized, suspended in PBS or Tris buffer38'