Each lot of this antibody is quality control tested by intracellular flow cytometry using our True-Phos? Perm Buffer in Cell Suspensions Protocol. For flow cytometric staining, the suggested use of this reagent is ≤ 0.5 ?g per million cells in 100 ?l volume. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 750? excites at 405 nm and emits at 750 nm. The bandpass filter 780/60 nm is recommended for detection, although filter optimization may be required depending on other fluorophores used. Be sure to verify that your cytometer configuration and software setup are appropriate for detecting this channel. Refer to your instrument manual or manufacturer for support. Brilliant Violet 750? is a trademark of Sirigen Group Ltd.

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.

1. Abrams J, et al. 1992. Immunol. Rev. 127:5. (Neut)
2. Abrams J, et al. 1995. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.20
3. Mo X, et al. 1995. J. Virol. 69:1288.
4. Sarawar S, et al. 1994. J. Immunol. 153:1246.
5. Via C, et al. 2001. J. Immunol. 167:6821. (Neut)
6. Infante-Duarte C, et al. 2000 J. Immunol. 165:6107. (FC)
7. Jacobs M, et al. 2000. Immunology 100:494. (IHC)
8. Marinova-Mutachieva L, et al. 1997. Clin. Exp. Immunol. 107:507. (IHC)
9. Williams RO, et al. 2000. J. Immunol. 165:7240. (IHC)
10. Scanga CA, et al. 1999. Infect. Immun. 67:4531. (Neut)
11. Akilov OE, et al. 2007. J. Leukoc. Biol. 2007;10.1189/jlb.0706439. (FC)
12. Lawson BR, et al. 2007. J. Immunol. 178:5366. (FC)
13. Patole PS, et al. 2005. J. Am. Soc. Nephrol. 16:3273. PubMed
14. Wu S, et al. 2005. Neurosci Lett. 394:158. PubMed
15. Carlson MJ, et al. 2009. Blood 113:1365. PubMed
16. Shivakumar P, et al. 2017. JCI Insight. 2:e88747 1. PubMed
17. Kearney CJ, et al. 2017. Cell Death Differ. 10.1038/cdd.2017.94. PubMed

1. Fitzgerald K, et al. Eds. 2001. The Cytokine FactsBook. Academic Press, San Diego.
2. Beutler B, et al. 1988. Annu. Rev. Biochem. 57:505.
3. Beutler B, et al. 1989. Annu. Rev. Immunol. 7:625.
4. Tracey K, et al. 1993. Crit. Care Med. 21:S415.