Alexa Fluor® 647 Mouse Anti-Human THEMIS
Clone Q13-1103 (RUO)
- Brand BD Pharmingen™
- Alternative Name TSEPA; THMS1; C6orf190; C6orf207; SPOT; GASP; GRB2-associated protein
- Vol. Per Test 5 µl
- Isotype Mouse IgG1, κ
- Reactivity Human (QC Testing)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Human THEMIS Recombinant Protein
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.