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.

The A20 antibody does not react with leukocytes or?mouse cells expressing the CD45.2 alloantigen. Additional reported applications (for relevant formats of this clone) include: immunoprecipitation³, in vitro blocking of B cell responses^1,2, immunohistochemical staining of frozen sections: OCT embedded⁷ and acetone-fixed^4-6 (direct immunofluorescence detection with fluorochrome conjugated A20 was used in (5) and (6)).

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. Yakura H, et al. 1983. J. Exp. Med. 157:1077. (Block)
2. Yakura H, et al. 1986. J. Immunol. 136:2729. (Block)
3. Shen FW, et al. 1986. Immunogenetics 24:146. (IP)
4. Suzuki K, et al. 2000. Immunity 13:691. (IHC-F)
5. Werner N, et al. 2002. Arterioscler. Thromb. Vasc. Biol. 22:1567. (IHC-F)
6. Lessner SM, et al. 2002. Am. J. Pathol. 160:2145. (FC, IHC-F)
7. Chen CC, et al. 2005. P. Natl. Acad. Sci. USA 102:11408 (IHC-F)
8. Pascal V, et al. 2007. J. Immunol. 179:1751. (FC)
9. Mende I, et al. 2006. Blood 107:1383. (IHC-F, FC)
10. Phan TG, et al. 2007. Nature Immunol. 8:992. (FC)
11. Wither DR, et al. 2009. J. Immunol. 183:5079. PubMed
12. Pascal V, et al.2007. J. Immunol. 179:1751. PubMed
13. Lee SW, et al. 2009. J. Immunol. 182:6753. PubMed
14. Takada K, et al. 2009. J. Exp Med. 206:2253. PubMed
15. Beamer CA, et al. 2007. Am. J. Respir. Cell. Mol. Biol. 37:729. (FC) PubMed
16. Li LX, et al. 2010. J. Immunol. 184:1728. PubMed
17. Hosoi A, et al. 2008. Cancer Res. 68:3941. (FC) PubMed
18. Kenna TJ, et al. 2008. Blood 111:2091. PubMed
19. Kohlmeier JE, et al. 2008. Immunity. 29:101. (FC) PubMed

All hematopoietic cells except mature erythrocytes and platelets of the CD45.1 strain of mice

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Trowbridge IS, et al. 1993. Annu. Rev. Immunol. 12:85.
3. Kishihara K, et al. 1993. Cell 74:143.
4. Pulido R, et al. 1988. J. Immunol. 140:3851.