Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤0.25 ?g per million cells in 100 ?l volume. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 421? excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421? is a trademark of Sirigen Group Ltd.

ELISA^1,3,4 or ELISPOT^2,6 Capture: The purified TRFK5 antibody is useful as a capture antibody for a sandwich ELISA or ELISPOT assay, when used in conjunction with biotinylated TRFK4 antibody (Cat. No. 504402) as the detection antibody for mouse IL-5, or used in conjunction with biotinylated JES1-5A10 antibody (Cat. No. 501002) as the detection antibody for human IL-5. The Ultra-LEAF? purified antibody is suggested for ELISPOT capture.

Flow Cytometry: The fluorochrome-labeled TRFK51 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify IL-5-producing cells within mixed cell populations.

Neutralization^1,9: The Ultra-LEAF? purified antibody (Endotoxin <0.1 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for neutralization of human IL-5 bioactivity (Cat. Nos. 504313-504318).

Additional reported applications (for the relevant formats) include: Western blotting, immunohistochemical staining of formalin-fixed paraffin-embedded tissue sections⁸ and paraformaldehyde-fixed, saponin-treated frozen tissue sections⁷, and immunocytochemistry.

Note: For testing human IL-5 in serum or plasma, BioLegend's ELISA Max? Sets (Cat. No. 430401 and 430404) are specially developed and recommended. For testing mouse IL-5 in serum, plasma or supernatant, BioLegend's ELISA MAX? Sets (Cat. Nos. 431201 and 431206) are specially developed and recommended.

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