Table 1.
Probes and antibodies used for the detection of SARS-CoV2 RNA and proteins and the concomitant host response.
| Reagent | Source/catalogue# | Dilution/pretreatment |
|---|---|---|
| SARS-CoV2 RNA probe | ACD/848561-C3 | RTU/AR and proteasea |
| ACE2 Ab | ProSci/3215 | 1:25,000/ARb |
| SARS-COV2 Spike S1 Ab | ProSci/9083 | 1:6000/AR |
| SARS-COV2 Spike S2 Ab | ProSci/9123 | 1:12,000/AR |
| SARS-COV2 envelope Ab | ProSci/3521 | 1:800/AR |
| SARS-COV2 matrix Ab | ProSci/3527 | 1:4000/AR |
| SARS-COV2 nucleocapsid Ab | ProSci/9099 | 1:13,000/AR |
| Caspase 3 Ab | Abcam/ab 4051 | 1:1200/AR |
| IL6 Ab | Abcam/ab6672 | 1: 7000/AR |
| TNFα Ab | Abcam/ab270264 | 1:5000/AR |
| Smooth muscle actin Ab | Ventana/790-2334 | RTU/AR |
| Complement component 6 | Proteintech/17239-1-AP | 1:1000/AR |
| CD11b | Abcam/ab 133657 | 1:7000/AR |
| CD163 | Abcam/182422 | 1:7000/AR |
| CD206 | Abcam/64693 | 1:6000/AR |
| CD68 | Ventana/790-2931 | RTU/AR |
| Type I collagen | Abcam/34710 | 1:600/AR |
| S-100 | Ventana/790-2914 | RTU/AR |
Each nonviral antibody was optimized in tonsil except for Complement component 6 which was optimized in COVID-19 lung with severe microangiopathy and pre-COVID normal lung.
Each viral antibody/probe was optimized in COVID-19 lung with high copy viral RNA in parallel with pre-COVID unremarkable lung.
a
As per ACD recommended protocol.
b
AR = antigen retrieval 30 min with Leica EDTA antigen retrieval solution; RTU = ready to use.