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?-A 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: immunohistochemical staining on frozen tissue^{4, 5},?formalin-fixed paraffin-embedded immunohistochemical staining⁶, and spatial biology (IBEX)^7,8.

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 CCTTGGCACCCGAGAATTCCA (PCR handle), and BAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA*A*A (capture sequence). B represents either 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. del C Salamone M, et al. 2001. J Leukoc Biol. 70:567.
2. de Fraissinette A, et al. 1988. Exp Hematol. 16:764.
3. Li D, et al. 2012. J Exp Med. 209:109. PubMed
4. Xu C, et al. 2010. Am J Hematol. 85:539 (IHC-F)
5. Gerlini G, et al. 2001. J Invest Dermatol. 117:576 (IHC-F)
6. Poposki J, et al. 2016. Clin Exp Allergy 45:384 (IHC-P) PubMed
7. Radtke AJ,?et al. 2020.?Proc Natl Acad Sci USA. 117:33455-33465. (SB)?PubMed
8. Radtke AJ,?et al. 2022.?Nat Protoc. 17:378-401. (SB)?PubMed

1. Cook CP, et al. 2022. Cell Rep Med. 3:100715. PubMed
2. Witkowski MT, et al. 2020. Cancer Cell. 37:867. PubMed
3. Hao Y, et al. 2021. Cell. 184:3573. PubMed

B cell subset, cortical thymocytes, dendritic cells, and Langerhans cells

1. Fainboim LM and del C. Salamone. 2002. J. Biol. Reg. Homeos. Ag. 16:125.
2. M. del Salamone C, et al. 2001. J. Leukocyte Biol. 70:567.
3. Zola H, et al. Eds. 2007. Leukocyte and Stromal Cell Molecules:The CD Markers. P42.