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: in vitro and in vivo blocking of ligand binding^1-5. For most successful immunofluorescent staining results, it may be important to maximize signal over background by using a relatively bright fluorochrome-antibody conjugate (Cat. No. 112706) or by using a high sensitivity, three-layer staining technique (e.g., including a biotinylated antibody (Cat. No. 112704) or biotinylated anti-rat IgG (Cat. No. 405402) second step, followed by SAv-PE (Cat. No. 405204)). The LEAF? purified antibody (Endotoxin < 0.1 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays. For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF? purified antibody (Cat. No. 112710) with a lower endotoxin limit than standard LEAF? purified antibodies (Endotoxin < 0.01 EU/?g).

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 NNNNNNNNNGCTTTAAGGCCGGTCCTAGC*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. O'Farrell A, et al. 1998. EMBO J. 17:1006. (Block)
2. Wang W, et al. 2004. FASEB J. 18:1043. (Block)
3. Vieira PL, et al. 2004. J. Immunol. 172:5986. (Block)
4. Iwashiro M, et al. 2001. P. Natl. Acad. Sci. USA 98:9226. (Block)
5. Vicari AP, et al. 2002. J. Exp. Med. 196:541. (Block)

Thymocytes, T and B cells, NK cells, monocytes, macrophages

1. Ho A, et al. 1993. P. Natl. Acad. Sci. USA 90:11267.
2. Tan JC, et al. 1993. J. Biol. Chem. 268:21053.
3. Niemand C, et al. 2003. J. Immunol. 170:3263.
4. Corinti S, et al. 2001. J. Immunol. 166:4312.