Table 1.
Reagents and conditions for immunohistochemistry and in situ hybridization
| Antibody/probe | Source* | Dilution | Pretreatment† | Predominant localization | Scoring scale, % |
||
|---|---|---|---|---|---|---|---|
| Negative | Weak | Strong | |||||
| HGAL | 115 | 1:20 | Citrate | Cytoplasmic | < 5 | > 5-30 | ≥ 30 |
| BCL6 | 2 | 1:40 | TRIS | Nuclear | < 5 | > 5-30 | ≥ 30 |
| CD10 | 3 | 1:40 | EDTA | Membrane | < 5 | > 5-30 | ≥ 30 |
| MUM1/IRF4 | 2 | 1:50 | EDTA | Nuclear | < 5 | > 5-30 | ≥ 30 |
| BCL2 | 2 | 1:1000 | TRIS | Cytoplasmic | < 20 | > 20-50 | ≥ 50 |
| Blimp1 | 418 | 1:10 | EDTA | Nuclear | < 5 | > 5-< 30 | ≥ 30 |
| pSTAT6 | 5 | 1:50 | Citrate | Nuclear | < 20 | ≥ 20 low‡ | ≥ 20 high‡ |
| EBV EBER | 6 | NA | NA | Nuclear | < 5 | > 5-30 | ≥ 30 |
| LMP2A | 719 | 1:100 | Citrate | Membrane | 0 | 1-10 | ≥ 10 |
For scoring scale, negative, weak, and strong staining is indicated numerically as 0, 2, and 3, respectively.
NA indicates not applicable.
Sources were as follows: 1, generated in our laboratory15; 2, DAKO; 3, Novocastra, New Castle-Upon-Tyne, United Kingdom; 4, kindly provided by Prof Katheryn Calame, Columbia University, New York, NY18; 5, Cell Signaling Technology, Danvers, MA; 6, INFORM EBER probe, Ventana; 7, kindly provided by Prof Gerald Niedobitek, Friedrich Alexander University, Erlangen, Germany.19
Pretreatment for heat-induced antigen retrieval consisted of microwaving with 1 of the following buffers: citrate (10 mM, pH 6.0, for 10 minutes), Tris (5 mM, pH 10.0, for 20 minutes), or EDTA (1 mM, pH 8.0, for 15 minutes).
pSTAT6 staining was scored based on low- and high-intensity nuclear labeling in at least 20% of Hodgkin/Reed-Sternberg cells.