FITC-I
Fluorescent Labeling Reagent
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Product codeF007 FITC-I
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CAS No.3326-32-7
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Chemical nameFluoresceiN-4-isothiocyanate
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MWC21H11NO5S=389.38
| Unit size | Price | Item Code |
|---|---|---|
| 100 mg | ||
| 500 mg |
Introduction
Fluoresceinisothiocyanate isomer-I (FITC, FTC) is a fluorescent reagent Fluorescein bonded with an amino group reactive NCS moiety, which is water-soluble and emits strong yellow-green fluorescence (λex=495 nm, λem=520 nm). Please store in a closed container in a cool place, protected from light and moisture. Since amino moiety easily reacts, it is suitable for fluorescent labeling of amino acids, peptides, proteins, enzymes, hormones, polysaccharides, bacteria, protozoa, fungi, viruses, and rickettsia. Therefore, it is used for diagnosis of collagen diseases, syphilis, and antinuclear antibodies induced by them by fluorescent antibody method.
Technical info
Solubility
100 mg/10 mL(Aceton)
References
References
1) H. Kawaguchi, K. Tuzimura, H. Maeda and N. Ishida, "Reaction of Fluorescein-isothiocyanate with Proteins and Amino Acids", J. Biochem., 1969, 66, 783.
2) H. Maeda, N. Ishida, H. Kawauchi and K. Tuzimura, "Reaction of Fluorescein-Isothiocyanate with Proteins and Amino Acids", J. Biochem., 1969, 65, 777.
3) W. B. Cherry, "Evaluation of Commercial Fluorescein Isothiocyanates Used in Fluorescent Antibody Studies", Stain Technology, 1969, 44, 179.
4) G. Radcliff, R. Waite, J. Lefevre, M. D. Poulik and D. M. Callewaert, "Quantification of Effector/Target Conjugation Involving Natural Killer(NK) or Lymphokine Activated Killer(LAK) Cells by Two-color Flow Cytometry", J. Immunol. Methods, 1991, 139, 281.
5) C. Souchier, M. Ffrench, M. Benchaib, R. Catallo and P. A. Bryon, "Methods for Cell Proloferation Analysis by Fluorescent Image Cytometry", Cytometry, 1995, 20, 203.
Q & A
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Q
How do we label proteins using FITC-I?
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A
An example of labeling by FITC-I is here attached.
This method is referred to the following book.
・G. T. Hermanson, ""Bioconjugate Techniques, Second Edition"", Elsevier, 2008.<Examples of protein labeling with FITC-I>
(1) Prepare a protein solution with a concentration of 2 mg/mL or higher in 0.1 mol/L carbonate buffer (pH 9.0).(2) Dissolve FITC-I in dry DMSO (1 mg/mL).
After preparation of FITC solution, wrap it in aluminum foil or otherwise protect it from light. *1(3) Add 50 to 100 μL of 1 mg/mL FITC solution to 1 mL of protein solution, and incubate at 4°C for at least 8 hours under shielded light. *2
(4) After reaction, elute with PBS using a gel filtration column such as Sephadex G-25 and purify it.
Handling and storage condition
| Appearance： | Yellowish orange powder |
|---|---|
| Purity (HPLC)： | ≧ 95.0 % |
| Solubility in Acetone： | To pass test (clear, yellow |
| Molar absorptivity： | ≧ 80,000 (around 492 nm) |
| IR spectrum： | Authentic |
| 0-5°C |
