Deep Dive: Fixing and Permeabilizing for Immunofluorescence
Type
Blog Post
...Antibodies
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Find plasmids encoding recombinant...targets, preventing your primary antibody from binding (Fig.1). The level of cross-linking that occurs...temperature, so you may be able to recover good antibody binding by tweaking those parameters, but in some cases...So, while organic solvents may improve antibody binding for some targets, they can destroy it for others... 1: Choice of fixation method impacts antibody binding. Formaldehyde cross-links the protein (red lines...results by not permeabilizing as you’ll prevent binding to any target protein that is still on the inside...reveal otherwise hidden epitopes. However, keep in mind that you will still contend with many of the same...