Application

Anti-Digoxigenin-Fluorescein, Fab fragments are used for the detection of digoxigenin-labeled compounds using:Digoxigenin-labeled sugars in glycoconjugate researchFluorescent in situ hybridization (FISH)ImmunohistocytochemistryIn situ hybridizationAnti-Digoxigenin-Fluorescein, Fab fragments cannot be used for membrane applications.

General description

Anti-Digoxigenin-Fluorescein, Fab fragments has fab fragments from polyclonal anti-digoxigenin antibodies, conjugated to fluorescein.

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 depends on application and substrate. The following concentrations should be taken as a guideline:Detection of digoxigenin-labeled sugars in glycoproteins: 50 to 20µg/ml Fluorescent in situ hybridization (FISH): 1 to 20µg/ml Immunohistocytochemistry: 20 to 50µg/ml In situ hybridization: 20 to 50µg/mlThe anti-DIG anitbody is highly specific and sensitive. Therefore, for FISH, a concentration of 1µg/ml is sufficient, and only in case of critical probes, this concentration may be raised to ﹤10µg/ml (with a distinct background to be expected).Working solution: ReconstitutionAdd 1 ml double-distilled water to a final concentration of 200µg/ml.Blocking:PBS, 0.5% bovine serum albumin (w/v), pH 7.4.1% 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. Furthermore, pH can be increased up to pH 8.5 to 9.0.Storage conditions (working solution): Always prepare fresh!

Reconstitution

Add 1 ml double-distilled water to a final concentration of 200 µg/ml.

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.