Will your antibodies work in IHC-F?

We do not routinely test our antibodies for IHC-F. However, we have found that antibodies recommended for IHC on paraffin embedded samples may work on fresh frozen samples as well. Because antibody performance in a given application is influenced by how the samples are prepared, antibodies that are known to work in immunocytochemistry (IF-IC) may be more likely to work in fresh frozen tissues, as the samples are handled more similarly.

When we do test our antibodies for IHC-F, we begin by optimizing the fixative for a given antibody, as we have seen over a number of comparisons that the choice of fixative can impact staining results. We generally compare the following fixatives: acetone, 10% neutral buffered formalin, 3-4% formaldehyde, and 3-4% formaldehyde followed by a methanol incubation. Once the fixative has been optimized we perform a small titration of the primary antibody to determine the optimal dilution.

Last updated: September 12, 2024