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.

H57-597 is a hamster mAb directed to an epitope of the C region of TCR ? chain¹². The H57-597 antibody does not cross-react with ?/d TCR-bearing T cells. Immobilized or soluble H57-597 antibody can activate a/? TCR-bearing T cells. Additional reported applications (for the relevant formats) for this antibody include: immunoprecipitation², in vitro stimulation^2,3, in vivo depletion^4-6, and immunohistochemical staining of acetone-fixed frozen sections^7,8,9. The Ultra-LEAF? purified antibody (Endotoxin <0.01 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. No. 109253-109258).

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. Gascoigne NJ. 1990. J. Biol. Chem. 265:9296.
2. Kruisbeek A, et al. 1991. In Current Protocols in Immunology. pp. 3.12.1. (Costim IP)
3. Davenport C, et al. 1995. J. Immunol. 155:3742. (Costim)
4. Drobyski W, et al. 1996. Blood 87:5355. (Deplete)
5. Kummer U, et al. 2001. Immunol. Lett. 75:153. (Deplete)
6. van der Heyde HC, et al. 1995. J. Immunol. 154:3985. (Deplete)
7. Tomita K, et al. 1999. Genes Dev. 13:1203. (IHC)
8. Podd BS, et al. 2006. J. Immunol. 176:6532. (IHC)
9. Ponomarev ED, et al. 2007. J. Immunol. 178:39. (IHC)
10. Chappaz S, et al. 2007. Blood doi:10.1182/blood-2007-02-074245. (FC) PubMed
11. Tsukumo S, et al. 2006. J.Immunol. 177:8365. (FC) PubMed
12. Grégoire C, et al. 1991. Proc. Natl. Acad. Sci USA 88:8077.

Majority of T cells and thymocytes (correlated to differentiation)

1. Davis MM, et al. 1998. Ann. Rev. Immunol. 16:523.
2. Huppa JB, et al. 2003. Nat. Immunol. 4:749.
3. Kubo R, et al. 1989. J. Immunol. 142:2736.