Table 2.
Overview of antibodies and chemicals used in the staining process
| RIG-I | EFTUD2 |
|---|---|
| Blocking solutiona: 20 min | Blocking solutionb: 5 min |
| Primary antibodyc | Primary antibodyd |
| 1:100 in ADe | 1:300 in PBSf |
| Incubation 1 h; 21 °C | Incubation 16,5 h; 4 °C |
| ImmPRESS polymer reagentg: 30 min | PostBlockh: 20 min |
| HRP Polymeri: 30 min | |
| Chromogen: DABj, 3 min | Chromogen: DABj, 2 min |
aNormal Horse Serum (NHS), 2.5%, ImmPRESS™ Anti-Mouse IgG Polymer Kit, VECTOR laboratories, Newark, United States; catalogue number: MP-7402
bZytoChem Plus HRP Polymer Kit (Mouse/Rabbit) 3 × 100, Zytomed Systems GmbH, Berlin, Germany; catalogue number POLHRP-100
cRIG-I-antibody: monoclonal IgG (mouse); LSBio, Seattle, United States; catalogue number: LS-C331000
dEFTUD2-antibody: polyclonal IgG (rabbit); Abcam, Cambridge, United Kingdom; catalogue number: ab72456
eAntibody Diluent, Agilent Technologies, Santa Clara, United States; catalogue number: S3022
fDulbecco ‘s Phosphate Buffered Saline, SigmaAlderich, Saint Louis, Missouri, United States
gAnti-Mouse IgG Polymer Kit, VECTOR laboratories, Newark, United States; catalogue number: MP-7402
hZytoChem Plus HRP Polymer Kit (Mouse/Rabbit); Zytomed Systems GmbH, Berlin, Germany
iVectastain Elite ABC Kit, diluted NORMAL serum; Vector laboratories, Newark, United States
jLiquid DAB + Substrate Chromogen System, REF K3468; Abcam, Cambridge, United Kingdom