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.

Additional reported applications (for the relevant formats) include: immunoprecipitation¹, in vitro costimulation of T and NK cells¹, in vitro blocking of allogeneic mixed leukocyte response and inhibition of MHC-unrestricted CTL cytotoxicity^3,4, in vitro induction of thymocyte differentiation^2,5-9,11, and immunohistochemical staining of acetone-fixed frozen sections. For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF? purified antibody (Endotoxin < 0.01 EU/?g, Azide-Free, 0.2 ?m filtered) (Cat. No. 102116).

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. Gross JA, et al. 1992. J. Immunol. 149:380. (IP, Costim)
2. Cibotti R, et al. 1997. Immunity 6:245. (Costim)
3. Masten BJ, et al. 1997. Am. J. Respir. Cell Mol. Biol. 16:335. (Block)
4. Nishio M, et al. 1996. J. Immunol. 157:4347. (Block)
5. Zhang N and He Y-W, 2005. J. Exp. Med. 202:395. (Costim)
6. Terrazas LI, et al. 2005. Intl. J. Parasitology. 35:1349. (Costim)
7. Perchonock CE, et al. 2006. Mol Cell Biol. 26(16):6005. (Costim)
8. Wang W, et al. 2007. J. Immunol. 178:4885. (Costim)
9. Pua HH, et al. 2007. J. Exp. Med. 204:25. (Costim)
10. Perchonock CE, et al. 2007. J. Immunol. 179:1768.
11. Barbi J, et al. 2007. Blood 110:2215.
12. Milpied P, et al. 2011. Blood 118:2993. PubMed
13. Cunningham NR, et al. 2011. Int Immunol. 23:693. PubMed
14. Crispin JC, et al. 2012. J. Immunol. 188:3567. PubMed
15. Li CR, et al. 2014. J Immunol. 192:1425. PubMed
16. Blankenhaus B, et al. 2014. PLoS Pathog. 10:1003913. PubMed

Thymocytes, CD4⁺, CD8⁺ peripheral T cells, NK cells

1. Barclay AN, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Lenschow DJ, et al. 1996. Annu. Rev. Immunol. 14:233.
3. Gross JA, et al. 1992. J. Immunol. 149:380.