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 RM4-5 antibody blocks the binding of GK1.5 antibody and H129.19 antibody to CD4⁺ T cells, but not RM4-4 antibody. Additional reported applications (for the relevant formats) include: blocking of ligand binding, in vivo depletion of CD4⁺ cells¹, and immunohistochemistry of acetone-fixed frozen tissue sections^2,3,11 and paraffin-embedded sections¹¹. Clone RM4-5 is not recommended for immunohistochemistry of formalin-fixed paraffin sections. Instead, acetone frozen or zinc-fixed paraffin sections are recommended. The Ultra-LEAF? Purified antibody (Endotoxin < 0.01 EU/?g, Azide-Free, 0.2 ?m filtered) is recommended for functional assays (Cat. No. 100575 and 100576).

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. Kruisbeek AM. 1991. In Curr. Protocols Immunol. pp. 4.1.1-4.1.5. (Block, Deplete)
2. Nitta H, et al. 1997. Cell Vision 4:73. (IHC)
3. Fan WY, et al. 2001. Exp. Biol. Med. 226:1045.
4. Muraille E, et al. 2003. Infect. Immun. 71:2704. (IHC)
5. León-Ponte M, et al. 2007. Blood 109:3139. (FC)
6. Bourdeau A, et al. 2007. Blood doi:10.1182/blood-2006-08-044370. (FC)
7. Matsumoto M, et al. 2007.J. Immunol.178:2499. PubMed
8. Shigeta A, et al. 2008. Blood 112:4915. PubMed
9. Zaborsky N, et al. 2010. J. Immunol. 184:725. PubMed
10. Rodrigues-Manzanet R, et al. 2010. P. Natl Acad Sci USA 107:8706. PubMed
11. Whiteland JL, et al. 1995. J. Histochem. Cytochem. 43:313. (IHC)

Majority of thymocytes, T cell subset

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Bierer BE, et al. 1989. Annu. Rev. Immunol. 7:579.
3. Janeway CA. 1992. Annu. Rev. Immunol. 10:645.