Each lot of this antibody is quality control tested by ELISA assay. For ELISA detection applications, a concentration range of 0.25 - 1.0 ?g/mL?is recommended. To obtain a linear standard curve, serial dilutions of TNF-α recombinant protein ranging from 500 to 4 pg/mL?are recommended for each ELISA plate.

For use as an ELISPOT detection antibody, a concentration range of 0.5 - 2.0 ?g/mL?is recommended. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 ?g per 10⁶ cells in 100 ?L?volume. It is recommended that the reagent be titrated for optimal performance for each application.

ELISA or ELISPOT Detection: The biotinylated MAb11 antibody is useful as the detection antibody in a sandwich ELISA or ELISPOT, when used in conjunction with the purified MAb1 antibody (Cat. No. 502802/502804) as the capture antibody.

Flow Cytometry^3,5,6,10: The fluorochrome-labeled MAb11 antibody is useful for intracellular and membrane-bound immunofluorescent staining and flow cytometric analysis to identify TNF-a-producing cells within mixed cell populations.

Additional reported applications (for the relevant formats) include: neutralization^1,2, immunohistochemical staining of paraformaldehyde-fixed, saponin-treated frozen tissue sections⁴ and acetone-fixed frozen tissue sections⁸, immunocytochemistry⁷, and immunofluorescence⁹. The MAb11 antibody can neutralize the bioactivity of natural or recombinant TNF-a.

Note: For testing human TNF-a in serum or plasma, BioLegend's ELISA Max? Sets (Cat. No. 430201 to 430206) are specially developed and recommended. The LEAF? purified antibody (Endotoxin <0.1 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for neutralization of human TNF-a bioactivity (Cat. No. 502922).

The Purified MAb1 antibody is useful in neutralization² and as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated MAb11 antibody (Cat. No. 502904/502914) as the detecting antibody.

Clone MAb11 cross-reacts to Cat¹¹

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