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 paraffin-embedded tissue sections¹¹, immunocytochemistry³, immunofluorescence microscopy^2,6, and blocking of CD4-independent infection by HIV-2 and CD4-dependent infection by some T cell-tropic isolates of HIV-1^4,5. Clone 12G5 may not be suitable for Western blotting.¹⁰ The Ultra-LEAF? purified antibody (Endotoxin <0.01 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. Nos. 306539 & 306540).

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. McKnight A, et al. 1997. J. Virol. 71:1692.
2. Endres MJ, et al. 1996. Cell 87:745. (Immunogen, IF)
3. Volin MV, et al. 1998. Biochem. Biophys. Res. Commun. 242:46. (ICC)
4. Berndt C, et al. 1998. P. Natl. Acad. Sci. USA 95:12556. (Block)
5. Ullrich CK, et al. 2000. Blood 96:1438. (Block)
6. Murga M, et al. 2005. Blood 105:1992. (IF)
7. Thompson BD. 2007. J. Biol. Chem. 282:9547. (FC) PubMed
8. Isnardi I, et al. 2010. Blood 115:5026. PubMed
9. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
10. Fischer T, et al. 2008. PLoS One 3:e4069.
11. Schmid BC, et al. 2004. Breast Cancer Res. Treat. 84:247. (IHC)

1. Bachireddy P, et al. 2021. Cell Rep. 37:109992. PubMed
2. Sokal A, et al. 2021. Cell. 184(5):1201-1213.e14. PubMed

T cells and B cells, dendritic cells, monocytes, granulocytes, hematopoietic progenitors, endothelial cells

1. Berger E, et al. 1999. Annu. Rev. Immunol. 17:657.
2. Loetscher P, et al. 2000. Adv. Immunol. 74:127.
3. Murphy P, et al. 2000. Pharmacol. Rev. 52:145.