Table 1.
Overview of the antibodies that allow immunostaining for NET components in paraffin-embedded tissue and the respective most effective antigen retrieval protocol.
| Source | Clone | Specificity | Host dilution | 37°C |
50°C |
50°C |
60°C |
96°C |
|||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Citrate pH6 | TRS pH9 | R-Univ pH7 | Citrate pH6 | TRS pH9 | Citrate pH6 | TRS pH9 | Citrate pH6 | TRS pH9 | |||||
| Abcam ab134211 | – | Histone H2B | ck 1:500 | H2B | ++1 N | +/− | ++1 N | ++1 N Q | ++1 N | +++ N | ++1 N | ++ N | +++ N |
| Antibodies-Online ABIN1735464 | – | Histone H3 | sh 1:100 | H3 | +/− | +/− N | ++1 N (Q) | ++1 N Q | ++1 N Q | ++1 N (Q) | ++ N | ++ N | ++1 N |
| Abcam ab5103 | – | Histone H3 citrulline (R2 + R8 + R17) | rb 1:50 | H3cit | ++1 (N) | +/− | ++ N Q | ++ N Q | ++1 (N) Q | +/− N | + (N) | +++ N | +++ N |
| Millipore 481001 | – | Neutrophil elastase | rb 1:50 | NE | ++ (N) | +/− | ++ N | ++ N | ++ (N) | + (N) | − | − | − |
| LS-B4244 | – | HN elanea | sh 1:200 | NEsh | ++ | ++ | ++ | ++ N | ++ N | ++ | ++ | − | − |
| R&D Systems AF3667 | – | Myeloperoxidase | gt 1:200 | MPO | ++ | ++ | ++ | ++ | ++ | ++ | ++ | ++ | ++ |
| Biorbyt orb316605 | – | CalgrA S100A8 MRP14 | rb 1:200 | CalA | +++ N | +++ N | +++ N | +++ N | +++ N | +++ N | +++ N | +++ N | +++ N |
| Biorbyt orb315186 | – | CalgrB S100A9 MRP14 | rb 1:200 | CalB | +++ N | +++ N | +++ N | +++ N | +++ N | +++ N | +++ N | +++ N | +++ N |
| BDPharmingen 551459 | 1A8 | Ly6G | rt 1:200 | Ly6G | ++ | +++ | +++ | +++ | +++ | +++ | +++ | +++ | +++ |
aOnly for human tissue.
R-Universal Buffer pH7 (RUB) 10× Aptum APO 0530500.
Target Retrieval Solution pH9 10mM Tris (TRS) 10× Dako S236784.
Target Retrieval Solution pH6 10mM Citrate 10× Dako S236984-3.
N, NET visualizations.
+++, strong staining of expected epitopes.
++, satisfying specific positive result.
+, moderate specific positive result.
+/−, weak or partially positive.
−, completely negative result.
Q, suitable as NET marker for quantification under mentioned conditions.
1, decondensed nuclei+++, nuclei +/−, or +.
(), limitations.