Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis and the oligomer sequence is confirmed by sequencing. TotalSeq?-C antibodies are compatible with 10x Genomics Chromium Single Cell Immune Profiling Solution.

To maximize performance, it is strongly recommended that the reagent be titrated for each application, and that you centrifuge the antibody dilution before adding to the cells at 14,000xg at 2 - 8°C for 10 minutes. Carefully pipette out the liquid avoiding the bottom of the tube and add to the cell suspension. For Proteogenomics analysis, the suggested starting amount of this reagent for titration is ≤ 1.0 ?g per million cells in 100 ?L volume. Refer to the corresponding TotalSeq? protocol for specific staining instructions.

While 1A8 recognizes only Ly-6G, clone RB6-8C5 recognizes both Ly-6G and Ly-6C. Clone RB6-8C5 binds with high affinity to mouse Ly-6G molecules and to a lower extent to Ly-6C¹⁵. Clone RB6-8C5 impairs the binding of anti-mouse Ly-6G clone 1A8¹⁵. However, clone RB6-8C5 is able to stain in the presence of anti-mouse Ly-6C clone HK1.4¹⁶.

Additional reported applications (for the relevant formats) include: immunohistochemistry⁹ of frozen sections¹⁰ and paraffin-embedded sections¹¹, depletion^{4, 12-14}, and spatial biology (IBEX)^20,21. The Ultra-LEAF? purified antibody (Endotoxin < 0.01 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for in vivo?studies or highly sensitive assays (Cat. No.?127632, 127649, 127650, 127661 and 127662).

TotalSeq? reagents are designed to profile protein levels at a single cell level following an optimized protocol similar to the CITE-seq workflow. A compatible single cell device (e.g. 10x Genomics Chromium System and Reagents) and sequencer (e.g. Illumina analyzers) are required. Please contact technical support for more information, or visit biolegend.com/totalseq.

The barcode flanking sequences are CGGAGATGTGTATAAGAGACAGNNNNNNNNNN (PCR handle), and NNNNNNNNNCCCATATAAGA*A*A (capture sequence). N represents either randomly selected A, C, G, or T, and * indicates a phosphorothioated bond, to prevent nuclease degradation.

View more applications data for this product in our Scientific Poster Library.

1. Fleming TJ, et al. 1993. J. Immunol. 151:2399. (FC)
2. Daley JM, et al. 2008. J. Leukocyte Biol. 83:1. (FC)
3. Dietlin TA, et al. 2007. J. Leukocyte Biol. 81:1205. (FC)
4. Daley J, et al. 2007. J. Leukocyte Biol. doi:10.1189. (Deplete) PubMed
5. Tadagavadi RK, et al. 2010. J. Immunol. 185:4904. PubMed
6. Sumagin R, et al. 2010. J. Immunol. 185:7057. PubMed
7. Guiducci C, et al. 2010. J. Exp Med. 207:2931. PubMed
8. Fujita M, et al. 2011. Cancer Res. 71:2664. PubMed
9. Van Leeuwen, et al. 2008. Arterioscler. Thromb. Vasc. Biol. 28:84. (IHC)
10. Kowanetz M, et al. 2010. P. Natl. Acad. Sci. USA 107:21248. [supplementary data] (IHC)
11. Esbona K, et al. 2016. Breast Cancer Res. 18:35. (IHC)
12. Wojtasiak M, et al. 2010. J. Gen. Virol. 91:2158. (FC, Deplete)
13. Jaeger BN, et al. 2012. J. Exp. Med. 209:565. (Deplete)
14. Wozniak KL, et al. 2012. BMC Immunol. 13:65 (FC, Deplete)
15. Ribechini E, et al. 2009. Eur. J. Immunol. 39:3538.
16. Ng LG, et al. 2011. J Invest. Dermatol. 131:2058. PubMed
17. Ma C, et al. 2012. J. Leukoc. Biol. 92:1199.
18. McCartney-Francis, N, et al. 2014. J Leukoc. Biol. 96:917. PubMed
19. Her Z, et al. 2014. EMBO Mol. Med. 7:24. PubMed
20. Radtke?AJ,?et al.?2020.?Proc Natl Acad Sci U S A.?117:33455-65. (SB)?PubMed
21. Radtke?AJ,?et al.?2022.?Nat Protoc.?17:378-401. (SB)?PubMed

1. Gomez S, et al. 2022. J Immunother Cancer. 10:. PubMed

Expressed on the majority of myeloid cells in bone marrow and peripheral granulocytes. The monoclonal antibody RB6-8C5 recognizes both Ly-6G and Ly-6C.

Fleming TJ, et al. 1993. J. Immunol. 151:2399.