Anti-THEMIS antibody

Cat.#: 176592

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Product Information

  • Product Name
    Anti-THEMIS antibody
  • Documents
  • Description
    Rabbit monoclonal antibody to THEMIS
  • Tested applications
    WB, ICC, IHC-P, FC
  • Species reactivity
    Human, Rat
  • Alternative names
    GASP antibody; SPOT antibody; TSEPA antibody; C6orf190 antibody; C6orf207 antibody
  • Isotype
    Rabbit IgG
  • Preparation
    This antigen of this antibody was synthetic peptide within c-terminal human themis.
  • Clonality
    Monoclonal
  • Formulation
    Liquid, 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
  • Storage instructions
    Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
  • Applications

    WB: 1:500-1:2,000

    ICC: 1:50-1:100

    IHC-P: 1:50-1:200

    FC: 1:50-1:100

  • Validations

    Fig1:; Western blot analysis of Themis on Jurkat cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody ( 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

    Fig1:; Western blot analysis of Themis on Jurkat cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody ( 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

    Fig2:; ICC staining of Themis in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody ( 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    Fig2:; ICC staining of Themis in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody ( 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    Fig3:; Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-Themis antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH; 2; O and PBS, and then probed with the primary antibody ( 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Fig3:; Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-Themis antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH; 2; O and PBS, and then probed with the primary antibody ( 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Fig4:; Immunohistochemical analysis of paraffin-embedded human appendix tissue using anti-Themis antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH; 2; O and PBS, and then probed with the primary antibody ( 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Fig4:; Immunohistochemical analysis of paraffin-embedded human appendix tissue using anti-Themis antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH; 2; O and PBS, and then probed with the primary antibody ( 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Fig5:; Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Themis antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH; 2; O and PBS, and then probed with the primary antibody ( 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Fig5:; Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Themis antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH; 2; O and PBS, and then probed with the primary antibody ( 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Fig6:; Flow cytometric analysis of Themis was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody ( 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Fig6:; Flow cytometric analysis of Themis was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody ( 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Background
  • References
    • Choi S. et. al. THEMIS: Two Models, Different Thresholds. Trends Immunol. 2017 Sep
    • Paster W. et. al. A THEMIS:SHP1 complex promotes T-cell survival. EMBO J. 2015 Feb

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"