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?-D antibodies are compatible with?Mission Bio’s Tapestri Single-Cell Sequencing Platform?for simultaneous detection of DNA and Protein.

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: blocking of ligand binding^1-3,?immunohistochemical staining of paraformaldehyde fixed frozen sections¹³, and spatial biology (IBEX)^15,16. The LEAF? purified antibody (Endotoxin <0.1 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. No. 329911 and 329912). For highly sensitive assays, we recommend Ultra-LEAF? purified antibody (Cat. No. 329926) with a lower endotoxin limit than standard LEAF? purified antibodies (Endotoxin <0.01 EU/?g).

TotalSeq?-D reagents are designed to profile protein expression at single cell level. The?Mission Bio Tapestri platform?and sequencer (e.g. Illumina analyzers) are required. Please contact?technical support?for more information, or visit?biolegend.com/totalseq/single-cell-dna

The barcode flanking sequences are CGAGATGACTACGCTACTCATGG (PCR handle), and GAGCCGATCTAGTATCTCAGT*C*G (capture sequence). * indicates a phosphorothioated bond, to prevent nuclease degradation.

View more applications data for this product in our?Application Technical Notes.

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11. Conrad J, et al. 2011. J. Immunol. 186:6871. PubMed
12. Salisch NC, et al. 2010. J. Immunol. 184:476. (Rhesus reactivity)
13. Li H and Pauza CD. 2015. Eur. J. Immunol. 45:298. (IHC)
14. Peterson VM, et al. 2017. Nat. Biotechnol. 35:936. (PG)
15. Radtke AJ,?et al. 2020.?Proc Natl Acad Sci USA. 117:33455-33465. (SB)?PubMed
16. Radtke AJ,?et al. 2022.?Nat Protoc. 17:378-401. (SB)?PubMed

Transiently expressed on CD4^- CD8^- thymocytes; upregulated in thymocytes and splenic T and B lymphocytes; expressed on activated myeloid cells