Immunofluorescence (IF/ICC) Troubleshooting: How to choose second antibody for IF?

There two points should be paid attention to when selecting the second antibody:
1. It is most important that select second antibody according to the source of the primary antibody. For example, if the primary antibody is raised in mouse, the second antibody should be select as goat-anti-mouse antibody, if the primary antibody is raised in rabbit, the second antibody should be select as goat-anti-rabbit antibody.
2.The fluorescent marker on second antibody is should selected carefully. The choice of fluorescent label is mainly based on the testing condition of the experiment test equipment and the requirements of the experiment itself.