Detects mouse Themis in direct ELISAs and Western blots. In Western blots, 100% cross-reactivity with recombinant human (rh) Themis (aa 2‑282) is observed under non-reducing conditions, and approximately 10% cross-reactivity with rhThemis (aa 2‑282) is observed under reducing conditions.
Monoclonal Rat IgG2B Clone # 719945
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant mouse Themis Lys122-Lys237 Accession # Q8BGW0
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Mouse Themis by Western Blot. Western blot shows lysates of mouse thymus tissue. PVDF membrane was probed with 1 µg/mL of Rat Anti-Mouse Themis Monoclonal Antibody (Catalog # MAB6816) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody. A specific band was detected for Themis at approximately 72 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Themis in Mouse Thymocytes by Flow Cytometry. Mouse thymocytes were stained with Rat Anti-Mouse Themis Monoclonal Antibody (Catalog # MAB6816, filled histogram) or isotype control antibody (Catalog # MAB0061, open histogram), followed by Phycoerythrin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0105B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
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 (thymocyte-expressed molecule involved in selection), also called Gasp (Grb2-associating protein), is a 72 kDa, 641 amino acid (aa) cytosolic phosphoprotein mainly expressed in late double-negative and double-positive thymocytes. It is involved in thymocyte positive and negative selection through regulation of TCR signaling. Within the region used as an immunogen, mouse Themis shares 87% and 96% aa sequence identity with human and rat 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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