Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining using the ?g size, the suggested use of this reagent is ≤1.0 ?g per million cells in 100 ?l volume. For immunofluorescent staining using the ?l size, the suggested use of this reagent is 5 ?l per million cells in 100 ?l staining volume or 5 ?l per 100 ?l of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 421? excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421? is a trademark of Sirigen Group Ltd.

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. Su W, et al. 2020. Cell Metabolism. 32(6):996-1011.e7. PubMed
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4. Kurniawan H, et al. 2020. Cell Metabolism. 31(5):920-936. PubMed
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7. Pasciuto E, et al. 2020. Cell. 182:625. PubMed
8. Tian M, et al. 2021. Elife. 10:. PubMed
9. Mondini M, et al. 2015. Mol Cancer Ther. 14:1336. PubMed
10. Hekim C, et al. 2017. Cancer Immunol Res. 5:157. PubMed
11. Phadke MS, et al. 2021. Cancer Immunol Res. 9:554. 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.