Anti-Digoxigenin-AP, Fab fragments

Description

• General description: Digoxigenin is a hapten, useful in labeling and detection of nucleic acids.[1] 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.
• 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
• Application: Use Anti-Digoxigenin-AP, Fab fragments for the detection of digoxigenin-labeled compounds using: cDNA arrayColony/plaque hybridizationDot blotELISAGel shift assay[2]Immunohistocytochemistry[3]In situ hybridization[4]Nonradioactive DNA sequencing blotNorthern blot[3]RNase protection assaySouthern blotWestern blotFluorescent in situ hybridization[5][6]Section in situ hybridization and whole mount in situ hybridization[7][8]Electrophoretic mobility shift assay[2]
• 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/mlELISA: 150 to 300 mU/mlImmunohistocytochemistry: 250 to 500 mU/mlIn situ hybridization: 1.5 to 7.5 U/mlSouthern blot: 150 mU/mlWestern 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!
• Analysis Note: Cross reactivity to digitoxin and digitoxigenin: <1 %No cross reactivity with other human estrogen or androgen steroids, e.g. estradiol or testosteroneCross reactivity with digoxin: not knownConjugate does not bind to itself at allNormally one molecule of the conjugate binds to one molecule digoxigenin, although ther are two possible binding sites for digoxigeninNonspecific binding to RNA is not expected
• Other Notes: For life science research only. Not for use in diagnostic procedures.