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11093274910 Roche

Anti-Digoxigenin-AP, Fab fragments

from sheep

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Properties

Related Categories Alphabetical Index, Antibodies, DE-DL, DIG System Antibodies, Molecular Biology,
conjugate   alkaline phosphatase conjugate
clone   polyclonal
biological source   sheep
form   solution
shipped in   wet ice
storage temp.   2-8°C
packaging   pkg of 200 μL (150 U)
antibody form   purified immunoglobulin
  purified immunoglobulin
isotype   IgG
Quality Level   100
antibody product type   primary antibodies
mfr. no.   Roche

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.

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 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:
• 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.

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis (COA)

Please Enter a Lot Number
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

Digoxigenin (DIG) Methods and Kits

The DIG System is the nonradioactive technology of choice to label and detect nucleic acids for multiple applications. The system is based on a steroid isolated from digitalis plants (Digitalis purpu...
Keywords: Amplification, Cloning, Diagnostic, Electrophoresis, Enzyme-linked immunosorbent assay, Gas chromatography, Gel electrophoresis, Immobilization, In Situ hybridization, Northern blot, Nucleic acid hybridization, Polymerase chain reaction, Purification, Transcription

Protocols

Anti-Digoxigenin-AP, Fab fragments Protocol & Troubleshooting

DIG Application Manual for Nonradioactive In Situ Hybridization 4th Edition, page 32, Correction for 50× Denhardt’s 50× Denhardt’s: 1% Ficoll (type 400), 1% polyvinylpyrrolidone, and 1% bovine serum ...
Keywords: In Situ hybridization, Nucleic acid hybridization, PAGE

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Peer-Reviewed Papers
15

References

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