FITC1
FluoroTag™ FITC Conjugation Kit
Sinônimo(s):
FITC
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About This Item
Código UNSPSC:
12352200
NACRES:
NA.32
Produtos recomendados
embalagem
pkg of (Kit contains reagents for 5 conjugations.)
Nível de qualidade
fluorescência
λex 495 nm; λem 525 nm
temperatura de armazenamento
2-8°C
Aplicação
The Flourotag FITC Conjugation Kit can be used for Immunohistochemisty and Immunocytochemistry using Flow Cytometry. It is also be used for the conjugation of FITC to peptide hormones, cytokines, growth factors and proteins.
Características e benefícios
- Suitable for both small (1 mg) and large (5 mg) scale conjugations
- Completely aqueous procedure - no DMF needed
- Fast gel filtration separation of conjugate from excess FITC
- Complete protocols for conjugation and F/P ratio determination
- Sufficient reagents for at least 5 conjugations of 5 mg protein each and for optimization of F/P ratio before scale-up
- References for applications and protocols
Princípio
Sigma offers a convenient kit for preparing FITC-labeled antibodies. Fluorescein isothiocyanate (FITC), Isomer 1, is a widely used fluorophore, popular because of its high quantum efficiency and stability when conjugated. FITC is yellow-orange in color with an absorption maximum at 495 nm. Upon excitation it emits a yellow-green color with an emission maximum at 525 nm. Conjugation occurs through free amino groups of proteins or peptides, forming a stable thiourea bond (see reaction). FITC conjugates of antibodies, lectins, hormones, and growth factors have been used in a variety of immunohistochemical and flow cytometry applications. The protocols have been optimized for antibodies, but may be adapted to other proteins by the end user.
Nota de análise
Procedure
1. Dissolve protein and FITC in carbonate-bicarbonate buffer.
2. Slowly add FITC to protein with stirring. Cover with foil and stir 2 hours at room temperature.
3. Separate conjugate from free FITC on G-25 column. Collect fractions.
4. Pool fractions containing conjugate.
5. Determine F/P ratio of conjugate spectrophotometrically.
6. Stabilize with 1% bovine serum albumin and 0.1% sodium azide and store at 0-5 °C.
1. Dissolve protein and FITC in carbonate-bicarbonate buffer.
2. Slowly add FITC to protein with stirring. Cover with foil and stir 2 hours at room temperature.
3. Separate conjugate from free FITC on G-25 column. Collect fractions.
4. Pool fractions containing conjugate.
5. Determine F/P ratio of conjugate spectrophotometrically.
6. Stabilize with 1% bovine serum albumin and 0.1% sodium azide and store at 0-5 °C.
Informações legais
FluoroTag is a trademark of Sigma-Aldrich Co. LLC
Somente componentes do kit
Nº do produto
Descrição
- Sephadex G-25 column, 3.5 mL 1
- Sephadex G-25 column, 9.1 mL 1
- Fluorescein isothiocyanate isomer I - F7250KC-2MG 2 mg
Componentes do kit também disponíveis separadamente
Nº do produto
Descrição
SDS
- P3813Phosphate buffered saline, powder, pH 7.4, for preparing 1 L solutions 5 pkgSDS
produto relacionado
Nº do produto
Descrição
Preços
Palavra indicadora
Warning
Frases de perigo
Declarações de precaução
Classificações de perigo
Eye Irrit. 2
Código de classe de armazenamento
10 - Combustible liquids
Ponto de fulgor (°F)
Not applicable
Ponto de fulgor (°C)
Not applicable
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