SBA Products [Fluorochrome Labeled Antibodies]

Preparation

Antibodies and F(ab')₂ fragments are conjugated to fluorescein isothiocyanate (FITC), tetramethylrhodamine isothiocyanate (TRITC) and Texas Red^TM (TXRD) under defined conditions, then purified by gel filtration and ion exchange chromatographies to select the most optimally conjugated fractions for use in immunofluorescence microscopy and flow cytometry.

Our SpectralRed^TM conjugate (SPRD) is a PE/CY5 tandem conjugate used as a third fluorochrome in multicolor flow cytometry applications when only a single 488 nm excitation source is available. Antibodies are covalently coupled to R-phycoerythrin (R-PE) and SpectralRed^TM (SPRD) using heterobifunctional linking reagents. The well-defined conditions employed for R-PE and SPRD labeling results in reagents optimally conjugated for flow cytometry applications.

Fluorescence Spectra

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Specificity

The specificity and performance of fluorochrome-labeled antibody are tested by particle concentration fluorescent immunoassay (PCFIA) and/or immunofluorescence staining with analysis by flow cytometry. To insure lot-to-lot consistency, each fluorochrome-labeled antibody is tested for conformance with characteristics of a standard reagent.
Working Dilutions

FITC-, TRITC- and TXRD-labeled antibodies fragments can be used routinely at less than or equal to 2µg/10⁶ cells, and R-PE conjugates at less than or equal to 1µg/10⁶ cells, when the staining analysis is by flow cytometry. For immunofluorescence microscopy applications, high conjugate concentrations may be required. The optimal staining reagent dilution for a particular immunofluorescence application should be determined by the investigator.

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Technical Assistance: * Toll Free US & Canada: 800-722-2255 * Telephone: 205-945-1774 * Fax: 205-945-8768 * Email

Form

FITC-, TRITC-, and TXRD-labeled antibodies are dialyzed into Dulbecco's phosphate buffered saline, pH 7.4, containing 0.1% sodium azide (PBS/NaN₃), passed through a 0.22 µ filter, diluted to stock concentration in PBS/NaN₃, and aseptically vialed. Each FITC-, TRITC- and TXRD-labeled product is supplied at a concentration of 0.5-1.0mg/mL in PBS/NaN₃.

R-PE and SPRD conjugates are dialyzed into PBS/NaN₃, filtered, diluted in stabilization buffer and aseptically vialed. R-PE- and SPRD-labeled product is supplied at a concentration of 0.1-0.25 mg/mL.
Storage and Stability

Recommended storage for FITC-, TRITC and TXRD-labeled products is at 2-8°C or in aliquots at -20°C. To increase long-term stability, glycerol (40%) may be added to the product prior to preparing aliquots and freezing at -20°C.

R-PE- and SPRD-labeled products contain a stabilizing reagent for storage at 2-8°C. R-PE and SPRD conjugates should not be stored at temperatures below 2°C - Do not freeze!

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Technical Assistance: * Toll Free US & Canada: 800-722-2255 * Telephone: 205-945-1774 * Fax: 205-945-8768 * Email

Applications

Flow cytometry
Immunofluorescence microscopy
Immunocytochemistry

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Technical Assistance: * Toll Free US & Canada: 800-722-2255 * Telephone: 205-945-1774 * Fax: 205-945-8768 * Email

Preparation
Antibodies and F(ab')₂ fragments are conjugated to fluorescein isothiocyanate (FITC), tetramethylrhodamine isothiocyanate (TRITC) and Texas Red^TM (TXRD) under defined conditions, then purified by gel filtration and ion exchange chromatographies to select the most optimally conjugated fractions for use in immunofluorescence microscopy and flow cytometry.		Our SpectralRed^TM conjugate (SPRD) is a PE/CY5 tandem conjugate used as a third fluorochrome in multicolor flow cytometry applications when only a single 488 nm excitation source is available. Antibodies are covalently coupled to R-phycoerythrin (R-PE) and SpectralRed^TM (SPRD) using heterobifunctional linking reagents. The well-defined conditions employed for R-PE and SPRD labeling results in reagents optimally conjugated for flow cytometry applications.