Each lot of this antibody is quality control tested by ELISA assay. For ELISA capture applications, a concentration range of 4-8 ?g/ml is recommended. To obtain a linear standard curve, serial dilutions of IL-10 recombinant protein ranging from 2000 to 15 pg/ml are recommended for each ELISA plate. It is recommended that the reagent be titrated for optimal performance for each application.
* For ELISA capture, it is very critical to use 0.2 M Sodium Phosphate Buffer, pH 6.5 (11.8g Na₂HPO₄, 16.1g NaH₂PO₄; q.s. to 1.0 L) as coating buffer.
Note: Carbonate buffer, pH 9.5 should not be used as coating buffer for JES5-2A5. It may cause high background and lower sensitivity.

ELISA or ELISPOT Capture^1-5,7,10: The purified JES5-2A5 antibody is useful as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated JES5-16E3 antibody (Cat. No. 505004) as the detecting antibody. The LEAF? purified antibody is suggested for ELISPOT capture.

ELISA or ELISPOT Detection: The biotinylated JES5-2A5 antibody is useful as the detecting antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the Purified JES5-16E3 antibody (Cat. No. 505002/505012) as the capture antibody.

Neutralization^2,8,9: The LEAF? purified antibody (Endotoxin <0.1 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for neutralization of mouse IL-10 bioactivity in vivo and in vitro (Cat. No. 504904).

Additional reported applications (for the relevant formats) include: Western blotting¹² and in vivo capture⁶.

Note: For testing mouse IL-10 in serum, plasma or supernatant, BioLegend's ELISA Max? Sets (Cat. No. 431401 to 431406) are specially developed and recommended.

1. Sander B, et al. 1993. J. Immunol. Meth. 166:201.
2. Abrams J, et al. 1992. Immunol. Rev. 127:5.
3. Abrams J, 1995. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.20.
4. Mo X, et al. 1995. J. Virol. 69:1288.
5. Sarawar S, et al. 1994. J. Immunol. 153:1246.
6. Finkelman F, et al. 2003. Curr. Prot. Immunol. John Wiley & Sons, New York. Unit 6.28.
7. Brummel R and Lenert P, 2005. J. Immunol. 174:2429.
8. Hara M, et al. 2001. J. Immunol. 166:3789.
9. Riemann M, et al. 2005. J. Immunol. 175:3560.
10. Xu G, et al. 2007. J. Immunol. 179:5358.
11. Guo Z, et al. 2013. J. Immunol. 190:4337. PubMed
12. van der Poll T, et al. 1995. J. Immunol. 155:5397. (WB)

1. Min CK, et al. 2018. Front Immunol. 1.779166667. PubMed
2. Maerz JK, et al. 2020. Front Immunol. 10:3093. PubMed
3. Campos–Mora M, et al. 2019. Front Immunol. 1.029166667. PubMed
4. Yan J, et al. 2020. Cell Rep. 107820:31. PubMed
5. Di Conza G, et al. 2021. Nat Immunol. 22:1403. PubMed
6. Ranjan K, et al. 2021. J Clin Invest. 131:. PubMed
7. Xu G, et al. 2007. J Immunol. 179:5358. PubMed
8. Riemann M, et al. 2005. J Immunol. 175:3560. PubMed
9. He L, et al. 2018. Immunology. 153:84. PubMed

1. Fitzgerald, K., et al. Eds. 2001. The Cytokine FactsBook. Academic Press, San Diego.
2. de Waal-Malefy, T, et al. 1992. Curr. Opin. Immunol. 4:314.
3. Howard, M., et al. 1992. Immunol. Today. 13:198.
4. Quesniaux, V. 1992. Research Immunol. 143:385.