The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/50 - 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Use buffer at pH 6 for antigen retrieval.
1/250 - 1/500.
Use a concentration of 1 - 4 µg/ml. The following steps are recommended prior to antibody incubation: cells fixation in ice cold 4% paraformaldehyde pH 7.2-7.3, followed by permeabilization with 0.1% Triton X-100 in PBS for 3x5 minutes.
FunctionSevers actin filaments and accelerates their polymerization and depolymerization.
Tissue specificityWidely expressed. In the kidney, expression is apparent in podocytes and some tubule cells.
Involvement in diseaseDefects in INF2 are the cause of focal segmental glomerulosclerosis type 5 (FSGS5) [MIM:613237]. A renal pathology defined by the presence of segmental sclerosis in glomeruli and resulting in proteinuria, reduced glomerular filtration rate and edema. Renal insufficiency often progresses to end-stage renal disease, a highly morbid state requiring either dialysis therapy or kidney transplantation.
DomainThe WH2 domain acts as the DAD (diaphanous autoregulatory) domain and binds to actin monomers. Regulated by autoinhibition due to intramolecular GBD-DAD binding. The severing activity is dependent on covalent attachment of the FH2 domain to the C-terminus.