Antigen retrieval and primary antibody type affect sensitivity but not specificity of CD117 immunohistochemistry

Abstract

Antigen retrieval and changing between the three antibodies tested affect sensitivity but not specificity of CD117 immunohistochemistry. Antigen retrieval does not produce false-positive CD117 immunostaining.

Specificity of DOG1 (K9 clone) and protein kinase C theta (clone 27) as immunohistochemical markers of gastrointestinal stromal tumour

“…However, it is now recognized that up to 5% of GISTs may lack CD117 immunopositivity 1,2 and that several potential differential diagnoses of GIST may also show CD117 immunopositivity, e.g. synovial sarcoma, Kaposi’s sarcoma 3 . Consequently, KIT / PDGFRA genotyping is increasingly regarded as a key diagnostic test for GIST and especially its CD117 negative subtype 4 .…”

“…Further, while the numbers of mesenchymal neoplasms tested by the American group were vast, 6,7 the use of TMAs meant that only a tiny portion of each neoplasm would have been immunostained and visualized. Therefore, the possibility of focal DOG1 immunopositivity amongst non‐GIST mesenchymal neoplasms (as is seen, for example, with CD117 3 ) cannot be completely excluded. In 2008, a Novocastra DOG1 monoclonal antibody (clone K9) was released for commercial use.…”

Fat-associated lymphoid cluster in Cyprinus carpio: Characterisation and its relation with peritoneal haemangiosarcoma

“…The slides were blocked for 60 min with blocking solution (0.25% albumin in PBS) and then incubated at 37 C for 60 min with primary antibodies (Abcam™). In the market, no specific antibodies for all of the fish surface markers assessed in this study exist; however, in the case of polyclonal antibodies, cross-reactivity between some vertebrates was demonstrated [4,12,13,51,57,67]. The polyclonal anti-c-kit antibodies possess variations in sensitivity, but do not stain cells other than those expressing c-kit [57,67].…”

“…In the market, no specific antibodies for all of the fish surface markers assessed in this study exist; however, in the case of polyclonal antibodies, cross-reactivity between some vertebrates was demonstrated [4,12,13,51,57,67]. The polyclonal anti-c-kit antibodies possess variations in sensitivity, but do not stain cells other than those expressing c-kit [57,67]. In this study, the polyclonal anti-c-kit (ab16832 polyclonal Ab produced in rabbit conjugate with FITC) was selected because the same and similar Ab were used in other fish species [4,51].…”

“…The slides were placed in a Copplin glass with a recovery solution (citrate buffer, pH 6.0, Invitrogen™ 1:20 dilution) and then placed in an autoclave (120 C/15 min). This procedure was necessary because the antigen retrieval generally increased the sensitivity but did not alter the specificity of immunoreactivity [67].…”