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Key Documents

343210

Sigma-Aldrich

FITC Labeling Kit

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About This Item

Código UNSPSC:
41116133

fabricante / nombre comercial

Calbiochem®

Nivel de calidad

condiciones de almacenamiento

OK to freeze
protect from light

temp. de almacenamiento

2-8°C

Descripción general

Provides means to quickly and simply label antibodies or other proteins with fluorescein. Direct labeling of the primary antibody eliminates the need for a secondary antibody, and results in a lower background and overall higher signal-to-noise ratio. Each kit is sufficient to perform at least five conjugations on proteins of 100 µg to 2.5 mg each.

Componentes

Fluorescein (five vials), Carbonate Buffer Concentrate, DMF Solvent, PBS Buffer Concentrate, and a user protocol.

Advertencia

Toxicity: Multiple Toxicity Values, refer to MSDS (O)

Principio

The Calbiochem FITC Labeling Kit provides a means of quickly and simply labeling antibodies or other proteins with fluorescein. FITC-antibody conjugates are useful in many applications, such as flow cytometry and immunocytochemistry. Other proteins such as steroids may also be labeled with FITC. Direct labeling of the primary antibody eliminates the need for a secondary antibody, and therefore results in a lower background and an overall higher signal to noise ratio.

Nota de preparación

Antibody/Protein Preparation: Add 10 ml of carbonate buffer concentrate to 990 ml of distilled water and dialyze antibody in buffer overnight.

Nota de análisis

1. Read the absorbance of the FITC-antibody conjugate at two wavelengths: 280 nm and 495 nm. Use the 1X PBS/0.1%NaN3 as the spectrophotometric blank. Optimal absorbance readings of the conjugated protein will be within the range of 0.2-1.2 ABS.

2. Using the two absorbance readings, calculate the F/P molar ratio of the FITC/Protein conjugate using the following equation:<div class="Bio_doc_image">Figure 1: F/P Molar Ration
</div>

3. There are 2 equations used for the determination of the concentration of the FITC-antibody conjugate. If the protein used for conjugation was IgG, use the following equation to estimate the concentration of the labeled antibody:<div class="Bio_doc_image"><p class="title">Figure 2: Concentration of FITC-IgG
</div>If the protein conjugated was not IgG, use the following equation to estimate the concentration of conjugated protein:<div class="Bio_doc_image">Figure 3: Concentration of FITC-Protein
</div>Where E0.1% is the A280 reading of a 1 mg/ml solution of the unconjugated protein, as measured in a cuvette of 1 cm path length.

Otras notas

Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.

Información legal

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Pictogramas

Health hazard

Palabra de señalización

Danger

Clasificaciones de peligro

Eye Irrit. 2 - Repr. 1B - Resp. Sens. 1 - Skin Sens. 1

Código de clase de almacenamiento

6.1C - Combustible, acute toxic Cat.3 / toxic compounds or compounds which causing chronic effects

Clase de riesgo para el agua (WGK)

WGK 3


Certificados de análisis (COA)

Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»

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