ANTI-FLUORESCEIN-POD

General description

Fab fragments from polyclonal anti-fluorescein antibodies, conjugated to horseradish peroxidase. The reagent is an anti-fluorescein antibody, Fab fragments from sheep, conjugated with horse-radishperoxidase (POD). After immunization with fluorescein the sheep IgG (immunoglobulin G) 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 with POD and stabilized in 60mM Tris-Hepes-buffer, 0.4% bovine immunoglobulin (w/v), 0.2% Germall II (w/v), pH 7.2.

Specificity

The polyclonal antibody reacts with free and bound fluorescein.

Application

Anti-Fluorescein-POD, Fab fragments are used for detection of fluorescein-labeled compounds in:

• Dot blot
• ELISA (enzyme-linked immunosorbent assay)
• Immunohistocytochemistry
• In situ hybridization
• Southern blot
• Western blot

Preparation Note

Working concentration: Working concentration of conjugate depends on application and substrate. The following concentrations should be taken as a guideline:

• Dot blot: 150 mU/ml
• ELISA: 50 to 150 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: 500 to 1000 mU/ml

Working solution: 100 mM Tris-HCl, 150 mM NaCl, pH 7.5. If necessary 1% Blocking reagent (w/v), dry milk powder, 1 to 5% heat inactivated fetal calf serum (v/v) or sheep normal serum can be used for reduction of unspecific binding.

Reconstitution

Add 1 ml double-distilled water to a final concentration of 150 U/ml.

Other Notes

For life science research only. Not for use in diagnostic procedures.