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11093274910
Roche
Anti-Digoxigenin-AP, Fab fragments
from sheep
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Synonym(s):
digoxigenin, anti-digoxigenin
Quality Level
biological source
sheep
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
shipped in
wet ice
storage temp.
2-8°C
Related Categories
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.
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
No data available
Flash Point(C)
No data available
Certificate of Analysis
Enter Lot Number to search for Certificate of Analysis (COA).
Certificate of Origin
Enter Lot Number to search for Certificate of Origin (COO).
Product Information Sheet
Quotes and Ordering
Use of In Situ Hybridization to Examine Gene Expression in the Embryonic, Neonatal, and Adult Urogenital System.
Rumballe B A, et al.
Kidney Development: Methods and Protocols, 223-239 (2012)
Exorhodopsin and melanopsin systems in the pineal complex and brain at early developmental stages of Atlantic halibut (Hippoglossus hippoglossus).
Eilertsen M, et al.
The Journal of Comparative Neurology, 522(18), 4003-4022 (2014)
In situ hybridization methods for mouse whole mounts and tissue sections with and without additional β-galactosidase staining.
Komatsu Y, et al.
Mouse molecular embryology : methods and protocols, 1-15 (2014)
The digoxigenin (DIG) system for non-radioactive labelling and detection of nucleic acids-an overview.
<BIG>Holtke HJ, et al.</BIG>
Cellular and molecular biology (Noisy-le-Grand, France), 41, 883-905 (1995)
Tara C Carlisle et al.
The Journal of comparative neurology, 522(4), 861-875 (2013-08-14)
Electrical synapses are expressed prominently in the developing and mature nervous systems. Unlike chemical synapses, little is known about the developmental role of electrical synapses, reflecting the limitations imposed by the lack of selective pharmacological blockers. At a molecular level
Articles
Digoxigenin (DIG) Labeling Methods
Digoxigenin (DIG) labeling methods and kits for DNA and RNA DIG probes, random primed DNA labeling, nick translation labeling, 5’ and 3’ oligonucleotide end-labeling.
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