Description: The 4S.B3 monoclonal antibody reacts with interferon-gamma (IFN gamma). Human IFN gamma is a 17 kDa factor produced by activated T and NK cells and is an anti-viral and anti-parasitic cytokine. IFN gamma in synergy with other cytokines, such as TNF alpha, inhibits proliferation of normal and transformed cells. Immunomodulatory effects of IFN gamma are exerted on a wide range of cell types expressing the high affinity receptors for IFN gamma. Glycosylation of IFN gamma does not affect its biological activity.

Applications Reported: This 4S.B3 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This 4S.B3 antibody has been pre-diluted and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol A: Two step protocol for intracellular (cytoplasmic) proteins" located at Flow Protocols. This may be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Brilliant Ultra Violet™ 737 (BUV737) is a tandem dye that emits at 732 nm and is intended for use on cytometers equipped with an ultraviolet (355 nm) laser. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) or Brilliant Stain Buffer (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Our internal testing suggests that Brilliant Ultra Violet™ 737 (BUV737) is compatible with short-term methanol-based fixation, but should not be stored in buffers containing methanol for longer than one hour.

Excitation: 355 nm; Emission: 732 nm; Laser: Ultraviolet Laser.

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