Table 1.

Antibody and immunoblot protocol details

Antibody TargetApproximately, kDaProtein/Lane Cortex, μgProtein/Lane Medulla, μgPrimary Antibody SupplierDilutionReference
Claudin-22315, 7.5NAThermoFisher (#32–5600)1:200012
Claudin-72315, 7.5NAThermoFisher (#34–9100)1:100012
ENaC-α
 Full length >10080, 4020, 10Loffing (Zurich)1:500013,14
 Cleaved > 30
ENaC-β 10060, 3020, 10Loffing (Zurich)1:15,00013,14
ENaC-γ
 Full length > 8060, 3020, 10Palmer (Cornell)1:100013,14
 Cleaved > 60
NCC15060, 30NAMcDonough1:50007
NCCpS7115015, 7.5NALoffing (Zurich)1:500015
NHE3 8315, 7.58, 4McDonough1:200016
NHE3pS552 835, 2.58, 4Santa Cruz (sc-53962)1:100017
NKCC216015, 7.58, 4DSHB (Iowa)1:600018
NKCC2-pT96T10116015, 7.58, 4Forbush (Yale)1:200019
Na,K-ATPase–α1 1001, 0.51, 0.5Kashgarian (Yale)1:20020
  • ENaC-α, epithelial Na+ channel α-subunit; ENaC-β, epithelial Na+ channel β-subunit; ENaC-γ, epithelial Na+ channel γ-subunit; NCC, Na+-Cl cotransporter; NCCpS71, phosphorylated form of Na+-Cl cotransporter (phosphorylated at S71); NHE3, Na+/H+ exchanger isoform 3; NHE3pS552, phosphorylated form of Na+/H+ exchanger isoform 3 (phosphorylated at S552); NKCC2, Na+-K+-2Cl cotransporter isoform 2; NKCC2-pT96T101, phosphorylated form of Na+-K+-2Cl cotransporter isoform 2 (phosphorylated at Thr 96 and Thr 101).