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.

* Alexa Fluor? 700 has a maximum emission of 719 nm when it is excited at 633 nm / 635 nm. Prior to using Alexa Fluor? 700 conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome.

ELISA or ELISPOT Detection:?The biotinylated MP6-XT22 antibody is useful as a detection antibody for a sandwich ELISA or ELISPOT assay, when used in conjunction with purified 6B8 antibody (Cat. Nos. 510802 & 510804) as the capture antibody.
ELISA Capture: The purified MP6-XT22 antibody is useful as the capture antibody in a sandwich ELISA when used in conjunction with the biotinylated Poly5160 antibody (Cat. No. 516003) as the detection antibody and recombinant mouse TNF-α (Cat. No. 575209) as the standard.
Flow Cytometry^6,11,12:The fluorochrome-labeled MP6-XT22 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify TNF-a-producing cells within mixed cell populations.
Neutralization^1,5,10,16,17:The MP6-XT22 antibody can neutralize the bioactivity of natural or recombinant TNF-α. The LEAF? purified antibody (Endotoxin < 0.1 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for neutralization of mouse TNF-α bioactivity in vivo and in vitro(Cat. No. 506310). For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF? purified antibody (Cat. No. 506332) with a lower endotoxin limit than standard LEAF? purified antibodies (Endotoxin < 0.01 EU/?g).
Additional reported applications (for the relevant formats) include: Western blotting, immunohistochemical staining of paraformaldehyde-fixed, saponin-treated frozen tissue sections^7-9 in vivo detection⁵, immunofluorescence, and immunocytochemistry.
Note: For testing mouse TNF-α in serum, plasma or supernatant, BioLegend's ELISA Max? Sets (Cat. No. 430901) are specially developed and recommended.

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