|IP||0.2-1 μL/mg of lysate|
**********Please Note: Optimal concentrations/dilutions should be determined by the end user.**********
Anti-FH mouse monoclonal antibody at 1:500 dilution
Lane A: Hela Whole Cell Lysate
Lane B: 293T Whole Cell LysateLysates/proteins at 30 μg per lane.
Goat Anti-Mouse IgG H&L (Dylight800) at 1/15000 dilution.Developed using the Odyssey technique.
Performed under reducing conditions.Predicted band size:55 kDa
Observed band size:47 kDa
FH was immunoprecipitated using:
Lane A:0.5 mg Hela Whole Cell Lysate
Lane B:0.5 mg 293T Whole Cell Lysate
Lane C:0.5 mg A431 Whole Cell Lysate0.5 µL anti-FH mouse monoclonal antibody and 15 μl of 50 % Protein G agarose.Primary antibody:
Anti-FH mouse monoclonal antibody,at 1:250 dilutionSecondary antibody:
Dylight 800-labeled antibody to Mouse IgG (H+L), at 1:7500 dilutionDeveloped using the odssey technique.
Performed under reducing conditions.Predicted band size: 55 kDa
Observed band size: 48 kDa
Fumarate Hydratase (FH) is an enzymatic component of the tricarboxylic acid (TCA) cycle, or Krebs cycle, and catalyzes the formation of L-malate from fumarate. It exists in both a cytosolic form and an N-terminal extended form, differing only in the translation start site used. The N-terminal extended form is targeted to the mitochondrion, where the removal of the extension generates the same form as in the cytoplasm. Fumarate Hydratase is similar to some thermostable class II fumarases and functions as a homotetramer. Mutations in this gene can cause fumarase deficiency and lead to progressive encephalopathy. Individuals with hemizygous germline fumarate hydratase (FH) mutations are predisposed to renal cancer. These tumors predominantly exhibit functional inactivation of the remaining wild-type allele, implicating FH inactivation as a tumor-promoting event.