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: immunoprecipitation^1,2, in vitro blocking of IL-2 binding to low- and high-affinity receptors^1-4, growth inhibition of IL-2-dependent T-cell lines^1-4, in vivo depletion of CD25⁺CD4⁺ Treg cells^5-8,10, and immunohistochemical staining of acetone-fixed frozen sections². PC61 antibody recognizes a different epitope than 3C7 antibody (Cat. No. 101902).?For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF? purified antibody (Cat. No. 102040) with endotoxin < 0.01 EU/?g, Azide-Free, 0.2 ?m filtered.?

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. Lowenthal JW, et al. 1985. Nature 315:669. (IP, Block)
2. Ceredig R, et al. 1985. Nature 314:98. (IP, IHC, Block)
3. Lowenthal JW, et al. 1985. J. Immunol. 135:3988. (Block)
4. Moreau JL, et al. 1987. Eur. J. Immunol. 17:929. (Block)
5. Takahashi T, et al. 2000. J. Exp. Med. 192:303. (Deplete)
6. Onizuka S, et al. 1999. Cancer Res. 59:3128. (Deplete)
7. Lei TC, et al. 2005. Blood 105:4865. (Deplete)
8. Pasare C, et al. 2004. Immunity 21:733. (Deplete)
9. León-Ponte M, et al. 2007. Blood 109:3139.
10. Cao OW, et al. 2007. Blood doi:10.1182/blood-2007-02-073304. (Deplete)
11. Benson MJ, et al. 2007. J. Exp. Med. doi:10.1084/jem.20070719.
12. Liu F, et al. 2011. Arch Toxicol. 85:1383. PubMed
13. Anguela XM, et al. 2013. Diabetes. 62:551. PubMed

Activated T cells and B cells, thymocyte subset, pre-B cells, T regulatory cells

1. Taniguchi T, et al. 1993. Cell 73:5-8.
2. Waldmann TA. 1991. J?Biol?Chem. 266:2681-4.
3. Read S, et al. 2000. J Exp?Med. 192:295-302.
4. Lowenthal JW, et al. 1985. J?Immunol. 135:3988-94.