FITC-conjugated antibodies are fluorescent markers used to identify proteins in various techniques. FITC absorbs energy at 495 nm and emits at 521 nm, allowing for systematic detection. FITC antibodies can be used directly or indirectly to label proteins in immunocytochemistry, enzyme and protein activity assays, flow cytometry, and Western blotting. They can be purchased from molecular biology companies or manufactured by research laboratories.
A fluorescein isothiocyanate (FITC) conjugated antibody is a highly specific fluorescent molecular marker for a particular protein or primary antibody. They are used to identify proteins in immunocytochemistry, enzyme and protein activity assays, flow cytometry, and Western blotting. Most FITC antibodies can be purchased through specialized molecular biology companies or manufactured by individual research laboratories.
FITC is a dye often conjugated or linked to an antibody to create a specific fluorescent marker. When exposed to electromagnetic energy at a specific wavelength, fluorochromes are chemicals that will absorb the energy and emit a photon at a specific wavelength. FITC absorbs energy at 495 nm and emits a photon at 521 nm. Each dye has a different absorption and emission range, allowing for the systematic detection of many dye-bound antibodies in a piece of tissue or sample. This is similar to labeling different parts of a cell with different colors and then using special glasses to look at only one color at a time.
Antibodies can be linked or conjugated to the FITC molecule to produce a FITC antibody. This FITC antibody can be used to directly or indirectly label a protein. When a FITC antibody directly labels a protein, it acts as a primary antibody or as an antibody in direct contact with the antigen. Indirect use of a FITC antibody occurs when it is used as a secondary antibody, by specifically binding to a primary antibody. When a FITC-conjugated antibody is used directly, the labeling procedure takes less time, but the labeling is not as strong. Indirect labeling requires an additional step, but the signal or label is amplified because several FITC-tagged secondary antibodies can attach to a single primary antibody.
There are many uses for the FITC antibody in cellular and molecular biology. This type of antibody can be used alone or in combination with other monoclonal or polyclonal antibodies to label specific proteins within cells in a technique known as immunocytochemistry. FITC antibodies can also be used in techniques such as enzyme and protein activity assays, flow cytometry, and Western blotting.
FITC-conjugated primary and secondary antibodies are widely available from many molecular biology companies. It is also possible for laboratories to prepare their own primary antibodies and then use a kit to conjugate FITC to them. Some companies will take lab-prepared primary antibodies and conjugate them to FITC dye for a lab. FITC-conjugated secondary antibodies are not typically manufactured by individual research laboratories and are usually purchased from companies that specialize in the production of secondary antibodies.
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