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: immunohistochemical staining of acetone-fixed frozen sections^4,6,7 and formalin-fixed paraffin-embedded sections¹¹, immunoprecipitation¹, activation of T cells^2,3,5, Western blotting⁹, and spatial biology (IBEX)^16,17. The LEAF? purified antibody (Endotoxin < 0.1 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. No. 300413, 300414, and 300432). For highly sensitive assays, we recommend Ultra-LEAF? purified antibody (Cat. No. 300437, 300438, 300465, 300466, 300473, 300474) with a lower endotoxin limit than standard LEAF? purified antibodies (Endotoxin < 0.01 EU/?g).

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. Salmeron A, et al. 1991. J. Immunol. 147:3047. (IP)
2. Graves J, et al. 1991. J. Immunol. 146:2102. (Activ)
3. Lafont V, et al. 2000. J. Biol. Chem. 275:19282. (Activ)
4. Ryschich E, et al. 2003. Tissue Antigens 62:48. (IHC)
5. Thompson AG, et al. 2004. J. Immunol. 173:1671. (Activ)
6. Sakkas LI, et al. 1998. Clin. Diagn. Lab. Immun. 5:430. (IHC)
7. Mack CL, et al. 2004. Pediatr. Res. 56:79. (IHC)
8. Thakral D, et al. 2008. J. Immunol. 180:7431. (FC) PubMed
9. Van Dongen JJM, et al. 1988. Blood 71:603. (WB)
10. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
11. Pollard, K. et al. 1987. J. Histochem. Cytochem. 35:1329. (IHC)
12. Luckashenak N, et al. 2013. J. Immunol. 190:27. PubMed
13. Laurent AJ, et al. 2014. PLoS One. 9:103683. PubMed
14. Li J, et al. 2015. Cancer Res. 75:508. PubMed
15. Stoeckius M, et al. 2017. Nat. Methods. 14:865-868. (PG)
16. Radtke AJ,?et al. 2020.?Proc Natl Acad Sci USA. 117:33455-33465. (SB)?PubMed
17. Radtke AJ,?et al. 2022.?Nat Protoc. 17:378-401. (SB)?PubMed

1. Lau D, et al. 2022. Nat Commun. 13:4053. PubMed
2. Bachireddy P, et al. 2021. Cell Rep. 37:109992. PubMed
3. Shangguan S, et al. 2021. Elife. 10:. PubMed

Mature T and NK T cells, thymocyte differentiation

1. Barclay N, et al. 1993. The Leucocyte FactsBook. Academic Press. San Diego.
2. Beverly P, et al. 1981. Eur. J. Immunol. 11:329.
3. Lanier L, et al. 1986. J. Immunol. 137:2501-2507.