Description
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.
Application
Use Anti-Digoxigenin-AP, Fab fragments for the detection of digoxigenin-labeled compounds using:
cDNA array
Colony/plaque hybridization
Dot blot
ELISA
Gel shift assay
Immunohistocytochemistry
In situ hybridization
Nonradioactive DNA sequencing blot
Northern blot
RNase protection assay
Southern blot
Western blot
Fluorescent in situ hybridization
Section in situ hybridization and whole mount in situ hybridization
Electrophoretic mobility shift assay
Biochem/physiol Actions
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
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/ml
Working solution: Northern blot, Southern blot
100 mM Maleic acid, 150 mM NaCl, pH 7.5.
Western blot
50 mM Tris-HCl, 150 mM NaCl, pH 7.5
other applications
100 mM Tris-HCl, 150 mM NaCl, pH 7.5
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.
Storage conditions (working solution): Diluted conjugate is stable at 2 to 8 °C for 12 hours.
Always prepare freshly!
Analysis Note
Cross reactivity to digitoxin and digitoxigenin: 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
Other Notes
For life science research only. Not for use in diagnostic procedures.
