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: immunoprecipitation^1,2, immunofluorescence microscopy³, immunocytochemistry², blocking of Fas induced apoptosis¹, and Western blotting¹¹. Fas Ligand is expressed at low density on activated cells. 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. 306407) or by using a high sensitivity, three-layer staining technique (e.g., including a biotinylated antibody (Cat. No. 306404) or biotinylated anti-mouse IgG second step (Cat. No. 405303), followed by SAv-PE (Cat. No. 405204)). In addition, applying matrix metalloproteinases (MMPS) inhibitor in the cell culture system will increase the FasL staining intensity. The Ultra-LEAF? purified antibody (Endotoxin < 0.01 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. No. 306415 and 306416).

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. Kayagaki N, et al. 1995. J. Exp. Med. 182:1777.
2. Herr I, et al. 2000. Cell Death Differ. 7:129. (WB)
3. Bossi G, et al. 1999. Nature Medicine 5:90.
4. Andreola G, et al. 2002. J. Exp. Med. 195:1303.
5. Strauss L, et al. 2009. J. Immunol. 182:1469. PubMed
6. Li JH, et al. 2009. Am J. Pathol. 175:1124. PubMed
7. Zhao Q, et al. 2011. Fitoterapia. 82:735. PubMed
8. Kruger K, et al. 2011. J. Appl. Physiol. 110:1226. PubMed
9. Qin G, et al. 2012. J. Infect. Dis. PubMed
10. Khalid M, et al. 2012. J. Virol. 86:4906. PubMed
11. Qin G, et al. 2012. J. Infect. Dis. 205:1646. PubMed
12. Shrestha B, et al. 2012. J. Virol. 86:8937. PubMed
13. Mooren FC, et al. 2012. J. Appl. Physiol.113:1082. PubMed
14. Robinet P, et al. 2014. J Immunol. 192:5332. PubMed
15. Wang Y, et al. 2014. J Endocrinol. 222:151. PubMed

Activated T cells, NK cells, testis, eye, neutrophils, Clara type II cells

1. Suda T, et al. 1997. J. Exp. Med. 12:204.
2. Kayagaki N, et al. 1995. J. Exp. Med. 182:1777.
3. Tanaka M, et al. 1995. EMBO J. 14:1129.