TABLE 1.
Antisera used for immunofluorescence.
| Namea | Hostb | Dilution | Source | Blocking bufferc | Durationd |
|---|---|---|---|---|---|
| Primary antisera | |||||
| Nv-NF-κB | R | 1:100 | OpenBiosystems (custom) | TBS, 5% NGS, 1% BSA, 0.2% Tx | 1.5 h at RT |
| Nv-IκB | GP | 1:200 | OpenBiosystems (custom) | TBS, 5% NGS, 1% BSA, 0.2% Tx | 1.5 h at RT |
| Secondary antisera | |||||
| vFITC-α-R | G | 1:160 | Sigma, cat. no. F9887 | TBS, 5% NGS, 1% BSA, 0.2% Tx | 1.5 h at 37 °C |
| TR-α-R | G | 1:80 | Vector Labs, cat. no. TI-1000 | TBS, 5% NGS, 1% BSA, 0.2% Tx | 1.5 h at 37 °C |
| FITC-α-GP | G | 1:80 | ThermoFisher, cat. no. PA1-28,675 | TBS, 5% NGS, 1% BSA, 0.2% Tx | 1 h at 37 °C |
a
Antisera conjugations: fluorescein isothiocyanate, FITC; Texas Red, TR. Secondary antisera were created against antibodies from the indicated species (e.g., FITC-linked anti-guinea pig is listed as FITC-α-GP).
b
Antisera host animal: goat, G; guinea pig, GP; rabbit, R.
c
Blocking buffer was used to block membranes/tissue and to dilute primary antisera. Abbreviations: normal goat serum, NGS; Tris-buffered saline, TBS; Triton X-100, Tx. All percentages are measured in (vol/vol) except BSA (wt/vol).
d
Blocking was usually performed overnight at 4 °C on a shaker. Duration of incubation with primary antisera. Room temperature, RT.