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.

The RPA-T8 antibody does not block the binding of HIT8a antibody to CD8a. Additional reported applications of this antibody (for the relevant formats) include: immunohistochemical staining of paraformaldehyde-fixed frozen sections³ and costimulation of T cell responses⁴. This clone was tested in-house and does not work on formalin fixed paraffin-embedded (FFPE) tissue. The Ultra-LEAF? purified antibody (Endotoxin <0.1 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. Nos. 301073 & 301074).

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. Knapp W, et al. Eds. 1989. Leucocyte Typing IV. Oxford University Press. New York.
2. Schlossman S, et al. Eds. 1995. Leucocyte Typing V. Oxford University Press. New York.
3. Mack CL, et al. 2004. Pediatr. Res. 56:79. (IHC)
4. Magidovich E, et al. 2007. P. Natl. Acad. Sci. USA 104:13022.
5. Thakarl D, et al. 2008.J. immunol. 180:7431. PubMed
6. Kmieciak M, et al. 2009. J. Transl. Med. 7:89. (FC) PubMed
7. Thakral D, et al. 2008. J. Immunol. 180:7431. (FC) PubMed
8. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
9. Rout N, et al. 2010. PLoS One 5:e9787. (FC)
10. Stoeckius M, et al. 2017. Nat. Methods. 14:865. (PG)

1. Bachireddy P, et al. 2021. Cell Rep. 37:109992. PubMed
2. Hiltensperger M, et al. 2021. Nat Immunol. 22:880. PubMed

Majority of thymocytes, T cell subset, NK cells

1. Barclay N, et al. 1993. The Leucocyte Antigen FactsBook. Academic Press Inc. San Diego.