Analysis Note

Cross reactivity to digitoxin and digitoxigenin: ﹤1 % No cross reactivity with other human estrogen or androgen steroids, e.g. estradiol or testosterone Cross reactivity with digoxin: not known Conjugate does not bind to itself at all Normally one molecule of the conjugate binds to one molecule digoxigenin, although ther are two possible binding sites for digoxigenin Nonspecific binding to RNA is not expected

Application

Use Anti-Digoxigenin-AP, Fab fragments for the detection of digoxigenin-labeled compounds using: cDNA arrayColony/plaque hybridizationDot blotELISAGel shift assayImmunohistocytochemistryIn situ hybridizationNonradioactive DNA sequencing blotNorthern blotRNase protection assaySouthern blotWestern blotFluorescent in situ hybridizationSection in situ hybridization and whole mount in situ hybridizationElectrophoretic mobility shift assay

General description

Digoxigenin is a hapten, useful in labeling and detection of nucleic acids. This product contains Fab fragments from polyclonal anti-digoxigenin antibodies, conjugated to alkaline phosphatase. Anti-Digoxigenin-AP, Fab fragments are useful for the detection of digoxigenin-labeled compounds.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Preparation Note

After immunization with digoxigenin, sheep IgG was purified by ion-exchange chromatography, and the specific IgG was isolated by immunosorption. The Fab fragments obtained by papain digestion were purified by gel filtration, conjugated to the specific label, and stabilized in buffer.Working concentration: Working concentration of conjugate will depend on the application and substrate. The following concentrations should be taken as a guideline: •Dot blot: 150 mU/ml•ELISA: 150 to 300 mU/ml•Immunohistocytochemistry: 250 to 500 mU/ml•In situ hybridization: 1.5 to 7.5 U/ml•Southern blot: 150 mU/ml•Western blot: 250 to 500 mU/mlWorking solution: Northern blot, Southern blot100 mM Maleic acid, 150 mM NaCl, pH 7.5.Western blot50 mM Tris-HCl, 150 mM NaCl, pH 7.5other applications100 mM Tris-HCl, 150 mM NaCl, pH 7.51% Blocking reagent (w/v), 1 to 5% heat inactivated fetal calf serum (v/v) or sheep normal serum can be used for reduction of unspecific binding.Storage conditions (working solution): Diluted conjugate is stable at 2 to 8 °C for 12 hours.Always prepare freshly!

Specificity

The polyclonal antibody from sheep is specific to digoxigenin and digoxin and shows no cross-reactivity with other steroids, such as human estrogens and androgens.Heat inactivation: yes