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BV421 Mouse Anti-Human THEMIS
BV421 Mouse Anti-Human THEMIS

Two-color flow cytometric analysis of THEMIS expressed in human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells were fixed and permeabilized with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) and BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885). The cells were then stained with PE Mouse Anti-Human CD3 antibody (Cat. No. 555333/561808/561809) and either BD Horizon™ BV421 Mouse IgG1 κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon™ BV421 Mouse Anti-Human THEMIS antibody (Cat. No. 563721; Right Panel). The two-color flow cytometric dot plots showing the correlated expression of THEMIS (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light-scattering characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSRII Flow Cytometer System.

Two-color flow cytometric analysis of THEMIS expressed in human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells were fixed and permeabilized with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) and BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885). The cells were then stained with PE Mouse Anti-Human CD3 antibody (Cat. No. 555333/561808/561809) and either BD Horizon™ BV421 Mouse IgG1 κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon™ BV421 Mouse Anti-Human THEMIS antibody (Cat. No. 563721; Right Panel). The two-color flow cytometric dot plots showing the correlated expression of THEMIS (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light-scattering characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSRII Flow Cytometer System.

Product Details
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BD Horizon™
TSEPA; THMS1; C6orf190; C6orf207; SPOT; GASP; GRB2-associated protein
Human (QC Testing)
Mouse IgG1, κ
Human THEMIS Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2738388
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. Brilliant Violet™ 421 is a trademark of Sirigen.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563721 Rev. 1
Antibody Details
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Q13-1103

The Q13-1103 monoclonal antibody specifically binds to human THEMIS (Thymocyte-expressed molecule involved in selection). The human THEMIS gene encodes a 73-kDa signaling protein that is also known as thymocyte selection pathway associated (TSEPA), signaling phosphoprotein specific for T cells (SPOT) and GRB2-associated protein (GASP). The homologous mouse protein is encoded by the Themis gene. THEMIS mRNA is detected in the thymus, lymph nodes, spleen but not in the bone marrow or nonlymphoid tissues. It is expressed by all thymocytes including double- (high levels) and single-positive αβ T cells, γδ T cells and peripheral T cells (low levels) but not by B cells and NK cells. THEMIS plays a central role in T cell development including positive and negative selection processes in the thymus. It is involved in TCR signaling and gets phosphorylated quickly after TCR stimulation. In the mouse system, Themis-deficient double-positive thymocytes reportedly expressed lower levels of genes involved in later stages of T cell development including those related to cellular metabolism, cell cycle and survival functions. Themis-deficient thymocytes showed impaired selection that lead to the production of fewer mature thymocytes. The Q13-1103 antibody crossreacted with mouse Themis in Western blot testing of cellular lysates but did not crossreact when tested by immunofluorescent staining and flow cytometric analysis of cells.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant Violet™ family of dyes.  With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421  can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421  conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

563721 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
563721 Rev.1
Citations & References
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Development References (4)

  1. Fu G, Vallée S, Rybakin V, et al. Themis controls thymocyte selection through regulation of T cell antigen receptor-mediated signaling. Nat Immunol. 2009; 10(8):848-856. (Biology). View Reference
  2. Johnson AL, Aravind L, Shulzhenko N, et al. Themis is a member of a new metazoan gene family and is required for the completion of thymocyte positive selection. Nat Immunol. 2009; 10(8):831-838. (Biology). View Reference
  3. Lesourne R, Uehara S, Lee J, et al. Themis, a T cell-specific protein important for late thymocyte development. Nat Immunol. 2009; 10(8):840-847. (Biology). View Reference
  4. Patrick MS, Oda H, Hayakawa K, et al. Gasp, a Grb2-associating protein, is critical for positive selection of thymocytes. Proc Natl Acad Sci U S A. 2009; 106(38):16345-16350. (Biology). View Reference
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563721 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.