11093274910
Roche
Anti-Digoxigenin-AP, Fab fragments
from sheep
Synonym(s):
anti-digoxigenin, digoxigenin
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About This Item
UNSPSC Code:
12352203
Recommended Products
biological source
sheep
Quality Level
conjugate
alkaline phosphatase conjugate
antibody form
purified immunoglobulin
antibody product type
primary antibodies
clone
polyclonal
form
solution
packaging
pkg of 200 μL (150 U)
manufacturer/tradename
Roche
isotype
IgG
storage temp.
2-8°C
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.
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
Heat inactivation: yes
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
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:
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!
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: <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
Other Notes
For life science research only. Not for use in diagnostic procedures.
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