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?-B antibodies are compatible with 10x Genomics Single Cell Gene Expression Solutions.

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: immunoprecipitation³, immunohistochemical staining of acetone-fixed frozen sections³, immunofluorescence microscopy^{5, 9} (Alexa Fluor? 488 conjugated N418 was used for IHC in frozen sections¹⁰), and spatial biology (IBEX)^22,23.

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 GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNNNNNNN (PCR handle), and NNNNNNNNNGCTTTAAGGCCGGTCCTAGC*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. Granucci F, et al. 1997. J. Immunol. 159:1794.
2. Stokes RW, et al. 1998. J. Immunol. 160:5514.
3. Metlay JP, et al. 1990. J. Exp. Med. 171:1753. (IHC, IP)
4. Ma XT, et al. 2006. Cancer Research 66:1169.
5. Chin RK, et al. 2006. J. Immunol. 177:290. (IF)
6. Cervantes-Barragan L, et al. 2007. Blood 109:1131. (FC) PubMed
7. Turnquist HR, et al. 2007. J. Immunol. 178:7018. (FC) PubMed
8. Benson MJ, et al. 2007. J. Exp. Med. doi:10.1084/jem.20070719. (FC) PubMed
9. You Y, et al. 2009. J. Immunol. 182:7343. (IF) PubMed
10. Roland CL, et al. 2009. Mol. Cancer Res. 8:1761. (IHC, FC) PubMed
11. Wikstrom M, et al.2006. J. Immunol. 177:913. PubMed
12. Pericolini E, et al. 2008. J. Leukocyte Biol. 83:1286. PubMed
13. Randall LM, et al. 2008. Infect. Immun.76:3312. PubMed
14. Fahlen-Yrild L, et al. 2009. J. Immunol. 183:5032. PubMed
15. Osterholzer JJ, et al. 2009. J. Immunol. 183:8044. PubMed
16. Bankoti J, et al. 2010. Toxicol. Sci. 115:422. (FC) PubMed
17. Eisenach PA, et al. 2010. J Cell Sci. 123:4182. PubMed
18. Leppin K, et al. 2014. Invest. Ophthalmol. Vis. Sci. 55:3603. PubMed
19. Sakai F, et al. 2014. PLoS One. 9:105370. PubMed
20. Gibbins JD, et al. 2014. Blood. 124:2953. PubMed
21. White CE, et al. 2015. J Immunol. 194:697. PubMed
22. Lu X, et al. 2015. J Immunol. 194:2011. PubMed
23. Radtke?AJ,?et al.?2020.?Proc Natl Acad Sci U S A.?117:33455-65. (SB)?PubMed
24. Radtke?AJ,?et al.?2022.?Nat Protoc.?17:378-401. (SB)?PubMed

Dendritic cells, NK cells, intestinal intraepithelial lymphocytes (IEL), some activated T cells

1. Barclay A, et al. 1997. The Leukocyte Antigen Facts Book Academic Press.
2. Springer TA. 1994. Cell 76:301.
3. Lopez-Rodriguez C, et al. 1996. J. Immunol. 156:3780.