Table 2.
Mouse Immunoblot Details
| Antibody target | ~kDa | μg/lane cortex | Primary ab supplier | Ab host | Dilution | Time | Secondary ab supplier | Host and target | Dilution | Time |
|---|---|---|---|---|---|---|---|---|---|---|
|
(Pro)renin receptor Full‐length cleaved |
~37 ~28 |
40 | Sigma | Rb | 1:1000 | O/N | Invitrogen | GAR 680 | 1:5000 | 1 h |
|
ENaC‐α Full‐length cleaved |
~80 ~20 |
40, 20 |
Loffing (Zurich) |
Rb | 1:5000 | 1 h | Invitrogen | GAR 680 | 1:5000 | 1 h |
| ENaC‐β | ~90 | 40, 20 |
Loffing (Zurich) |
Rb | 1:1500 | O/N | Invitrogen | GAR 680 | 1:5000 | 1 h |
|
ENaC‐γ Full‐length cleaved |
~80 ~60 |
40, 20 |
Loffing (Zurich) |
Rb | 1:1000 | O/N | Invitrogen | GAR 680 | 1:5000 | 1 h |
| NHE3 | ~83 | 40, 20 | McDonough | Rb | 1:2000 | O/N | Invitrogen | GAR 680 | 1:5000 | 1 h |
| NHE3pS552 | ~83 | 5, 2.5 |
Santa Cruz (sc‐53 962) |
Mu | 1:1000 | 2 h | Invitrogen | GAM 680 | 1:5000 | 1 h |
| NKCC2 | 160 | 20, 10 |
DSHB (Iowa) |
Mu | 1:6000 | O/N | LI‐COR | GAM 800 | 1:5000 | 1 h |
| NKCC2p‐ S87 | 160 | 40, 20 |
DSTT (Dundee) |
Sh | 1:2500 | 2 h | Invitrogen | DAS 680 | 1:5000 | 1 h |
Ab indicates antibody; ENAC, epithelial sodium channel; Mu, mouse; NHE3, sodium‐hydrogen antiporter 3; NKCC2, Na‐K‐Cl cotransporter 2; O/N, overnight; Rb, rabbit; and Sh, sheep.