Rabbit Anti-Fluorescein Isothiocyanate (FITC) (HRP)
FITC (fluorescein isothiocyanate) is a fluorochrome dye that absorbs ultraviolet or blue light causing molecules to become excited and emit a visible yellow-green light. This emission ceases upon removal of the light causing the excitation. Fluorochrome labeling provides rapid, accurate localization of antigen-antibody interaction when one of the reactants is part of a cell, tissue or other biological structure. FITC is a commonly used marker for antibodies in immunofluorescent techniques since the conjugation of FITC to proteins is relatively easy and does not, in general, destroy the biological activity of the labeled protein. FITC is widely used as a hapten to label different proteins.
Applications
Suitable for use in Western Blot, Immunohistochemistry and ELISA. Other applications have not been tested.
Recommended Dilutions
ELISA: 1:200-1:1000 Western Blot: 1:20-1:200 Immunohistochemistry (frozen): 1:20-1:200 Optimal dilutions to be determined by the researcher.
Substitution Ratio
(Estimated) HRP:IgG is >1.5
Storage and Stability
Store product at 4°C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20°C. Aliquots are stable at -20°C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Note: Sodium azide is a potent inhibitor of peroxidase and should not be added to HRP conjugates. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Form
Supplied as a liquid in PBS, pH 7.2, 1% BSA, 0.0095% MIT. Sodium azide is a potent inhibitor of peroxidase and should not be added to HRP conjugates..
Purity
Purified by affinity chromatography.
Specificity
Recognizes Fluorescein. No crossreactivity with immunoglobulins of any species. The isothiocyanate group reacts with amino terminal and primary amines in proteins.
