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Abcam's FITC Conjugation Kit provides a simple and quick process to conjugate your primary antibodies with FITC. FITC is a green fluorescent dye which has an excitation wavelength of 495nm and an emission wavelength of 519nm.
The conjugated antibody can be used straight away in WB, ELISA, IHC, Immunofluorescence (ICC/IF), Flow cytometry and FRET etc
Learn more about buffer compatibility, protein/secondary antibody conjugation and labelling chemistry in our FAQs.
The antibody to be labelled should be purified, in an appropriate buffer for conjugation and at a suitable concentration, as described in the protocol booklet. If not, consider using our antibody purification and concentration kits.
The purified antibody to be labeled should ideally be in 10-50mM amine-free buffer (e.g. MES, MOPS, HEPES, PBS), pH range 6.5 to 8.5. If the buffer is more concentrated or outside this pH range contact the technical support team.
Common non-buffering salts (e.g. sodium chloride), chelating agents (e.g. EDTA), and sugars have no effect on conjugation efficiency. Azide (0.02 to 0.1%) and BSA (0.1 to 0.5%) have little or no effect. Glycerol up to 50% has no effect.
Avoid buffer components that are nucleophilic, as these may react with the kit chemicals. Primary amines (e.g. amino acids or ethanolamine) and thiols (e.g. mercaptoethanol or DTT) fall within this class (Note: Tris-based buffers should be avoided). If your buffer contains primary amines and/or thiols, you should consider using our Concentration and Purification Kits (ab102778 or ab102784).
Recommended amount and volume of antibody for optimal results
|Pack size||Amount of antibody||Volume of antibody|
|Up to 20 µg||10-20 µg||4-10 µl|
|Up to 200 µg||100-200 µg||40-100 µl|
|Up to 2 mg||1-2 mg||400-1000 µl|
Antibody concentrations of 1-4 mg/ml generally give optimal results. For conditions outside the suggested ranges contact the technical support team.
Storage of conjugates
Storage at 4°C is recommended for any conjugate. A preservative may be desirable for long-term storage. Other storage conditions may also be satisfactory. The best conditions for any particular conjugate must be determined by experimentation.
Labeling of the antibody will not work if the conjugation blocks the active paratope. This situation is rare.
|Components||300 µg||30 µg||1 mg|
|FITC mix||3 x 100µg||3 vials||1 vial|
|Modifier reagent||1 vial||1 vial||1 vial|
|Quencher reagent||1 vial||1 vial||1 vial|
Our Abpromise guarantee covers the use of ab102884 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Conjugation||Use at an assay dependent dilution.|
This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.