Water-soluble organic solvent improves paraffin displacement in all-in-one epitope retrieval buffers

Abstract

An all-in-one epitope retrieval buffer was developed to solubilize molten paraffin. This study illustrates that common all-in-one buffer additives such as surfactants and wetting agents do not solubilize molten paraffin but a water-soluble organic solvent (WSOS) does. This study also shows that the WSOS does not negatively impact immunohistochemical staining reactions. The addition of a WSOS into an all-in-one epitope retrieval buffer formulation maintains paraffin in solution when slides mounted with tissue sections are removed from the buffer. This action is an advantage for buffer tanks in which slides are in a vertical orientation; the slides are not recoated with molten paraffin.

When Tissue Antigens and Antibodies Get Along

“…148 The advantage of these solutions is obvious, but reported instances of incomplete paraffin removal could affect the outcome of the IHC. 114 This is more commonly observed with solutions based on surfactants and wetting agents, which tend to incompletely solubilize the molten paraffin. The addition of a water-soluble organic agent appears to solve this problem without negatively affecting the IHC reaction.…”

“…The addition of a water-soluble organic agent appears to solve this problem without negatively affecting the IHC reaction. 114 …”

Protocol to quantify immune cell distribution from the vasculature to the glioma microenvironment on sequential immunofluorescence multiplex images

“… Note: The reagents are based on patented technology and are common practice reagents in immunohistochemistry experiments. 8 , 9 , 10 , 11 Validate all antibodies using conventional immunohistochemistry and/or IF staining on positive control tissue in conjunction with the corresponding fluorophore and the spectral 4′,6-diamidino-2-pheynlindole (DAPI; Thermo Fisher Scientific) counterstain. For optimal concentration and the best signal/noise ratio, test the antibodies at three different dilutions, starting with the manufacturer-recommended dilution (MRD), then MRD/2 and MRD/4.…”

“… Note: The reagents are based on patented technology and are common practice reagents in immunohistochemistry experiments. 8 , 9 , 10 , 11 …”