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 ≤0.5 ?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 570? excites at 405 nm and emits at 570 nm. The bandpass filter 585/42 nm is recommended for detection, although filter optimization may be required depending on other fluorophores used. Be sure to verify that your cytometer configuration and software setup are appropriate for detecting this channel. Refer to your instrument manual or manufacturer for support. Brilliant Violet 570? is a trademark of Sirigen Group Ltd.

The RM4-5 antibody blocks the binding of GK1.5 antibody and H129.19 antibody to CD4⁺ T cells, but not RM4-4 antibody. Additional reported applications (for the relevant formats) include: blocking of ligand binding, in vivo depletion of CD4⁺ cells¹, and immunohistochemistry of acetone-fixed frozen tissue sections^2,3,11 and paraffin-embedded sections¹¹. Clone RM4-5 is not recommended for immunohistochemistry of formalin-fixed paraffin sections. Instead, acetone frozen or zinc-fixed paraffin sections are recommended. The Ultra-LEAF? Purified antibody (Endotoxin < 0.01 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. No. 100575 and 100576).

1. Kruisbeek AM. 1991. In Curr. Protocols Immunol. pp. 4.1.1-4.1.5. (Block, Deplete)
2. Nitta H, et al. 1997. Cell Vision 4:73. (IHC)
3. Fan WY, et al. 2001. Exp. Biol. Med. 226:1045.
4. Muraille E, et al. 2003. Infect. Immun. 71:2704. (IHC)
5. León-Ponte M, et al. 2007. Blood 109:3139. (FC)
6. Bourdeau A, et al. 2007. Blood doi:10.1182/blood-2006-08-044370. (FC)
7. Matsumoto M, et al. 2007.J. Immunol.178:2499. PubMed
8. Shigeta A, et al. 2008. Blood 112:4915. PubMed
9. Zaborsky N, et al. 2010. J. Immunol. 184:725. PubMed
10. Rodrigues-Manzanet R, et al. 2010. P. Natl Acad Sci USA 107:8706. PubMed
11. Whiteland JL, et al. 1995. J. Histochem. Cytochem. 43:313. (IHC)

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Majority of thymocytes, T cell subset

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