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?-B 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: Western blotting¹, in vitro induction of thymocyte maturation², complement-mediated cytotoxicity³, and immunohistochemistry of acetone-fixed frozen sections⁴, formalin-fixed paraffin-embedded sections⁵ and zinc-fixed paraffin-embedded sections¹⁰. The Ultra-LEAF? purified antibody (Endotoxin < 0.01 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. No. 101845 and 101846).

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 GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNNNNNNN (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. Springer T, et al. 1978. Eur. J. Immunol. 8:539. (WB)
2. Crowley M, et al. 1989. Cell. Immunol. 118:108. (FA)
3. Veillette A, et al. 1989. J. Exp. Med. 170:1671. (FA)
4. Pandelakis A Flavell RA 1999 JEM 189:855 (FC, IHC)
5. Liu JQ, et al. 2007 J. Immunol. 178:6227. (FC, IF)
6. Chappaz S, et al. 2007. Blood doi:10.1182/blood-2007-02-074245. (FC) PubMed
7. Rucci F, et al. 2010. Proc Natl Acad Sci USA. 107:3024. (FC) PubMed
8. Teague TK, et al. 2010. Int Immunol. 22:387. (FC) PubMed
9. Gracz AD, et al. 2010. Am J. Physiol Gastrointest Liver Physiol. 298:590. (FC) PubMed
10. Chen CY, et al. 2008. Endocrinology. 10:1210. (FC, IHC) PubMed
11. Qui Q, et al. 2010. J. Immunol. 184:1681. (FC) PubMed

B cells, granulyocytes, epithelial cells, thymocytes, monocytes, erythrocytes, dendritic cells

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Aigner S, et al. 1997. Blood 89:3385.
3. Hough MR, et al. 1996. J. Immunol. 156:479.
4. Liu Y, et al. 1992. J. Exp. Med. 175:437.