Antigen retrieval with heat

To localize ER[3 in frozen sections, antigen retrieval with heating is not required. 210-180-C050 antibodies (Alexis Corporation, Nottingham, UK) are obtained by immunizing rabbits with synthetic peptides representing the C-terminal amino acid residues 467-485 of human estrogen. 

Naito I, Ninomiya Y, Nomura S. Immunohistochemical diagnosis of Alport s syndrome in paraffin-embedded renal sections antigen retrieval with autoclave heating. Med. Electron Microsc. 2003 36 1-7. 

Rait VK, Xu L, O Leary TJ, et al. Modeling formalin fixation and antigen retrieval with bovine pancreatic RBase A II. Interrelationship of cross-linking, immuno-reactivity, and heat treatment. Lab. Invest. 2004 84 300-306. 

Ino H. Application of antigen retrieval by heating for double-label fluorescent immunohistochemistry with identical species-derived primary antibodies. J. Histochem. Cytochem. 2004 52 1219-1230. 

Figure 7.6 Peptides do not denature after baking (dry heat) or deparaffinization and antigen retrieval (wet heat). Peptide-coupled slides were treated as indicated on the Y-axis and then immunohistochemically stained. In this particular example, an ER peptide with an ER MAb was used. The resulting peptide spot intensity (mean pixel intensity on a 1-256 scale) was measured and is shown on the y-axis.The data represent the means and SD or triplicate measurements. The experiments on the left (solid bars) and the right (hatched bars) were conducted at different times and have no connection to one another. Adapted with permission from Sompuram et al.6...

Immunohistochemical detection of wild-type and mutant p53 proteins can be carried out in fresh-frozen as well as formalin-fixed and paraffin-embedded tissues. This is best accomplished by using antigen retrieval with microwave heating or other types of heating such as autoclaving. A number of monoclonal antibodies are commercially available, the characteristics and sources of which are listed in Tables 10.3 and 10.4 and on pages 50-51. 

Saito N, Konishi K, Takeda H, et al. Antigen retrieval trial for post-embedding immunoelectron microscopy by heating with several unmasking solutions. J. Histochem. Cytochem. 2003 51 989-994. 

Namimatsu S, Ghazizadeh M, Sugisaki Y. Reversing the effects of formalin fixation with citraconic anhydride and heat a universal antigen retrieval method. J. Histochem. Cytochem. 2005 53 3-11. 

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