Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is =1.0 ?g per million cells in 100 ?l volume. It is recommended that the reagent be titrated for optimal performance for each application.

* PerCP/Cyanine5.5 has a maximum absorption of 482 nm and a maximum emission of 690 nm.

The UC10-4B9 antibody can enhance T cell co-stimulation by blocking CTLA-4 interactions with the B7 co-receptors, favoring CD28 interactions. Additional reported applications (for the relevant formats) include: immunoprecipitation¹, in vitro stimulation, in vitro and in vivo blocking^1-4 of ligand binding, and as ELISA capture antibody⁵. To reduce non-specific binding to cells bearing Fc-receptors, pre-incubation of cells with anti-mouse CD16/CD32, clone 93 (Cat. No. 101301/101302), is recommended prior to immunofluorescent staining. For most successful immunofluorescent staining results, it may be important to maximize signal over background by using a relatively bright fluorochrome-antibody conjugate (Cat. No. 106306) or by using a high sensitivity, three-layer staining technique (e.g., including a biotinylated anti-Armenian hamster IgG (Cat. No. 405501) second step, followed by SAv-PE (Cat. No. 405204)). The Ultra LEAF? purified antibody (Endotoxin < 0.01 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. No. 106327).

1. Walunas TL, et al. 1994. Immunity 1:405. (Block, IP)
2. Cilio CM, et al. 1998. J. Exp. Med. 188:1239. (Block)
3. Issazadeh S, et al. 1999. J. Immunol. 162:761. (Block)
4. McCoy K, et al. 1997. J. Exp. Med. 186:183. (Block)
5. Hsu HC, et al. 2007. J. Immunol. 178:5357. (ELISA Capture)
6. Sugita S, et al. 2010. Invest. Ophthalmol. Vis. Sci. 51:5783. PubMed

1. Liang Y, et al. 2020. J Immunol. 204:2984. PubMed
2. Russell SL, et al. 2019. Cell Rep. 29:3564. PubMed
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5. Webb ER, et al. 2022. iScience. 25:104995. PubMed
6. Hartmann W, et al. 2020. Cell Reports. 29(8):2243-2256.e4.. PubMed
7. Siolas D, et al. 2021. Cell Reports. 36(8):109578. PubMed

Activated T cells and B cells

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Allison JP, et al. 1995. Science 270:932.
3. Waterhouse P, et al. 1995. Science 270:985.
4. Linsley PS, et al. 1991. J. Exp. Med. 174:561.