Each lot of this antibody is quality control tested by ELISA assay. For ELISA capture applications, a concentration range of 1-4 ?g/ml is recommended. To obtain a linear standard curve, serial dilutions of mouse IL-5 recombinant protein ranging from 250 to 1 pg/ml are recommended for each ELISA plate when paired with TRFK4 (Cat. # 504402). To obtain a linear standard curve, serial dilutions of human IL-5 recombinant protein ranging from 250 to 1 pg/ml are recommended for each ELISA plate when paired with JES1-5A10 (Cat. # 501002). It is recommended that the reagent be titrated for optimal performance for each application.

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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