Each lot of this antibody is quality control tested by ELISA assay. For ELISA applications, a concentration range of 0.5 – 2.0 ?g/mL?is recommended. To obtain a linear standard curve, serial dilutions of mouse IL-6 recombinant protein ranging from 500 to 8 pg/mL?are recommended for each ELISA plate. It is recommended that the reagent be titrated for optimal performance for each application.

This product is suitable for use with the?Maxpar? Metal Labeling Kits.?For metal labeling using Maxpar? Ready antibodies,?proceed directly to the step to Partially Reduce the Antibody by adding 100 ?l of Maxpar? Ready antibody to 100 ?l of 4 mM TCEP-R in a 50 kDa filter and continue with the protocol. Always refer to the latest version of Maxpar? User Guide when conjugating Maxpar? Ready antibodies.

ELISA Capture^1-3,11 or ELISPOT Capture^4,5: The purified MP5-20F3 antibody is useful as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated MP5-32C11 antibody (Cat. No. 504602) as the detecting antibody and recombinant mouse IL-6 (Cat. No. 575709) as the standard. The Ultra-LEAF? purified antibody is suggested for ELISPOT capture.

Flow Cytometry: The fluorochrome-labeled MP5-20F3 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify IL-6 -producing cells within mixed cell populations. For intracellular cytokine staining protocol, please visit www.biolegend.com and click on the support section.

Neutralization^1-2,8-10: The Ultra-LEAF? purified antibody (Endotoxin <0.1 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for neutralization of mouse IL-6 bioactivity in vivo and in vitro (Cat. No. 504506).

Additional reported applications (for relevant formats of this clone) include: In Vivo Capture⁷, and immunohistochemical staining (paraformaldehyde-fixed/saponin-treated⁶, and acetone-fixed frozen tissue sections^{13, 14}).

Note: For testing mouse IL-6 in serum, plasma or supernatant, BioLegend's ELISA Max? Sets (Cat. No. 431301 to 431306) are specially developed and recommended.

Maxpar? is a registered trademark of Standard BioTools Inc.

1. Abrams J, et al. 1992. Immunol. Rev. 127:5. (ELISA Capture, Neut)
2. Sander B, et al. 1993. J. Immunol. Meth. 166:201. (ELISA Capture, Neut)
3. Abrams J, et al. 1995. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.20. (ELISA Capture)
4. Fujihashi K, et al. 1991. J. Immunol. Meth. 160:181. (ELISPOT Capture)
5. Klinman D, et al. 1994. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.19. (ELISPOT Capture)
6. Litton M, et al. 1994. J. Immunol Meth. 175:47. (IHC)
7. Finkelman F, et al. 2003. Curr. Prot. Immunol. John Wiley & Sons, New York. Unit 6.28. (In Vivo Capture)
8. Starnes HF Jr, et al. 1990. J. Immunol. 145:4185. (Neut)
9. Riedemann NC, et al. 2003. J. Immunol. 170:503. (Neut)
10. Rochman I, et al. 2005. J. Immunol. 174:4761. (Neut)
11. O'Connell PJ, et al. 2006. Blood 107:1010. (ELISA Capture)
12. Ozeki Y, et al. 2010. Int. Immunol. 22:179. PubMed
13. Galie M, et al. 2005. Carcinogenesis 26:1868 (IHC)
14. Schone MP, et al. 2000. J Immunol 165:6583 (IHC)
15. Keswani T, et al. 2014. Exp Parasitol. 141:82. PubMed
16. Notley CA, et al. 2015. J Immunol. 194:1621. PubMed

1. Fitzgerald K, et al. Eds. 2001. The Cytokine FactsBook. Academic Press, San Diego.
2. Hirano T. 1998. Intl. Rev. Immunol. 16:249.
3. Patterson P. 1992. Curr. Opin. Neurobiol. 2:94.
4. Van Oers M, et al. 1993. Ann. Hematology 66:219.