Detection of Mouse and Rat Themis by Western Blot.
Western blot shows lysates of mouse thymus tissue and Nb2‑11 rat lymphoma cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Mouse/Rat Themis Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6816) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Themis at approximately 73 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Themis (THymus-Expressed Molecule Involved in Selection; also C6orf190, Gasp and Thylex) is a 72-74 kDa member of the Themis family of proteins. It has restricted expression, being found in double negative #2-4 thymocytes, double positive thymocytes, and CD4+ plus CD8+ thymocytes and T cells. The molecule is known to constitute part of the TCR signalosome, along with LAT, Grb2 and PLC gamma 1, and is required for ERK and AP-1 activation. Themis appears to contribute to the process of positive selection. In particular, it is needed to prevent cell death during the selection process. Mouse Themis is 636 amino acids (aa) in length. It contains two globular CABIT (Cys-containing, all-beta in Themis) domains (aa 1-260 and 261-520) plus an NLS (aa 555-563). There are four isoform variants. One shows a 25 aa substitution for aa 238-636, a second possesses an alternative start site at Met90 coupled to a 13 aa substitution for aa 589-636, while third and fourth variants contain 7 aa and 17 aa substitutions for aa 589-636. Over aa 122-237, mouse Themis shares 96% and 87% aa identity with rat and human Themis, respectively.
Thymocyte-expressed Molecule Involved in Selection
Entrez Gene IDs:
387357 (Human); 210757 (Mouse)
bA325O24.3; bA325O24.4; C6orf190; C6orf207; FLJ40584; Gasp; GRB2-associated protein; MGC163388; protein THEMIS; signaling phosphoprotein specific for T cells; SPOT; Themis; thymocyte selection associated; thymocyte selection pathway associated; thymocyte-expressed molecule involved in selection; TSEPA
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