Each lot of this antibody is quality control tested by immunohistochemistry. For immunohistochemistry on frozen tissue sections, a concentration range of 2.5-5 μg/ml is suggested. For flow cytometric staining, the suggested use of this reagent is ≤0.25 ?g per million cells in 100 ?l volume.?It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor? 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.

Additional reported applications (for the relevant formats) include: blocking of CD4⁺ T cell activation^1,4,11, thymocyte costimulation³, in vitro and in vivo depletion^2,5-8, blocking of egg-sperm cell adhesion^1,4, immunohistochemical staining of acetone-fixed frozen sections^9,10, immunoprecipitation^1,2, and spatial biology (IBEX)^12,13. The GK1.5 antibody is able to block CD4 mediated cell adhesion and T cell activation.?Binding of GK1.5 antibody to CD4 T cells can be blocked by RM4-5 antibody, but not RM4-4 antibody. For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF? purified antibody (Cat. No. 100442) with a lower endotoxin limit than standard LEAF? purified antibodies (Endotoxin < 0.01 EU/?g).

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. Dialynas DP, et al. 1983. J. Immunol. 131:2445. (Block, IP)
2. Dialynas DP, et al. 1983. Immunol. Rev. 74:29. (IP, Deplete)
3. Wu L, et al. 1991. J. Exp. Med. 174:1617. (Costim)
4. Godfrey DI, et al. 1994. J. Immunol. 152:4783. (Block)
5. Gavett SH, et al. 1994. Am. J. Respir. Cell. Mol. Biol. 10:587. (Deplete)
6. Schuyler M, et al. 1994. Am. J. Respir. Crit. Care Med. 149:1286. (Deplete)
7. Ghobrial RR, et al. 1989. Clin. Immunol. Immunopathol. 52:486. (Deplete)
8. Israelski DM, et al. 1989. J. Immunol. 142:954. (Deplete)
9. Zheng B, et al. 1996. J. Exp. Med. 184:1083. (IHC)
10. Frei K, et al. 1997. J. Exp. Med. 185:2177. (IHC)
11. Felix NJ, et al. 2007. Nat. Immunol. 8:388. (Block)
12. Radtke?AJ,?et al.?2020.?Proc Natl Acad Sci U S A.?117:33455-65. (SB)?PubMed
13. Radtke?AJ,?et al.?2022.?Nat Protoc.?17:378-401. (SB)?PubMed

1. Wang D, et al. 2018. Immunity. 48:659. PubMed
2. Baptista AP et al. 2019. Immunity. 50(5):1188-1201 . PubMed
3. Wang B, et al. 2022. Nat Commun. 13:3821. PubMed
4. Ji Y, et al. 2017. Mucosal Immunol. 10.1038/mi.2016.119. PubMed
5. Bosnjak B, et al. 2019. Front Immunol. 10:840. PubMed
6. Kovacs SB, et al. 2021. STAR Protoc. 2:100244. PubMed
7. Tanaka Y, et al. 2017. J Immunol. 199:4016. PubMed
8. Kovacs SB, et al. 2020. Cell Reports. 32(4):107967. PubMed
9. Liu Q, et al. 2021. Adv Mater. 33:e2102852. PubMed
10. Chmielewski M and Abken H 2017. Cell Rep.. 10.1016/j.celrep.2017.11.063. PubMed
11. Wang X, et al. 2021. Sci Transl Med. 13:. PubMed

Majority of thymocytes, T cell subset

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Bierer BE, et al. 1989. Annu. Rev. Immunol. 7:579.
3. Janeway CA. 1992. Annu. Rev. Immunol. 10:645.