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 ≤ 0.25 ?g per million cells in 100 ?l volume.?For immunohistochemistry, a concentration range of 5.0 - 10.0 ?g/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor? 488 has a maximum emission of 519 nm when it is excited at 488 nm.

Additional reported applications (for the relevant formats) include: immunohistochemical staining^2,3,5-7 of acetone-fixed frozen sections, blocking^4,5,8 of ligand binding in vitro and in vivo,?immunoprecipitation¹?, and spatial biology (IBEX)^11,12. The Ultra-LEAF? purified antibody (Endotoxin < 0.01 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. No. 105727 & 105728).

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

1. Kinashi T, et al. 1995. J. Leukoc. Biol. 57:168. (IP)
2. Koni PA, et al. 2001. J. Exp. Med. 193:741. (IHC)
3. Ishiyama N, et al. 1998. Pathobiology 66:274. (IHC)
4. Kinashi T, et al. 1994. Blood Cells 20:25. (Block)
5. Baron JL, et al. 1994. J. Clin. Invest. 93:1700. (Block IHC)
6. Buck CA, et al. 1996. Cell Adhes. Commun. 4:69. (IHC)
7. Hata H, et al. 2004. J. Clin. Invest. 114:582. (IHC)
8. Meunier MC, et al. 2005. Nature Medicine 11:1222. (Block) PubMed
9. Monnier J, et al. 2012. J. Immunol. 189:956. PubMed
10. Motohashi N, et al. 2013. J Cell Sci. 126:2678. PubMed
11. Radtke?AJ,?et al.?2020.?Proc Natl Acad Sci U S A.?117:33455-65. (SB)?PubMed
12. Radtke?AJ,?et al.?2022.?Nat Protoc.?17:378-401. (SB)?PubMed

1. Ulyanova T, Phelps S, Papayannopoulou T 2016. Blood. 128: 1756 - 1765. PubMed
2. Baryawno N et al. 2019. Cell. 177(7):1915-1932 . PubMed
3. Severe N et al. 2019. Cell Stem Cell. 25(4):570-583 . PubMed

Bone marrow stromal cells, myeloid progenitors, splenic dendritic cells, activated endothelial cells

1. Barclay AN, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Kinashi T, et al. 1995. J. Leukoc. Biol. 57:168.
3. Bevilacquea MP. 1993. Annu. Rev. Immunol. 11:767.
4. Koni PA, et al. 2001. J. Exp. Med. 193:741.