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Invitrogen™ Themis Monoclonal Antibody (1TMYS), eFluor™ 660, eBioscience™
Description
This 1TMYS monoclonal antibody reacts with mouse Thymocyte-expressed molecule involved in selection (Themis), also known as Grb2-associating protein (Gasp). Themis is a cytoplasmic protein that is essential for positive selection and the development of single-positive thymocytes. Highest levels of expression are found in CD4+CD8+ double-positive thymocytes, with lower expression found in mature T cells. In addition to Grb1, Themis has been found to associate with PLC gamma 1 and LAT and is quickly phosphorylated by SLP-76 following TCR signaling. There are 5 isoforms of mouse Themis that are produced as a result of alternative splicing. The 1TMYS antibody recognizes isoforms 1, 2, 3, and 4, but does not recognize isoform 5. Applications Reported: This 1TMYS antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This 1TMYS antibody has been tested by intracellular staining and flow cytometric analysis of mouse thymocytes or splenocytes using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to BestProtocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins. This can be used at less than or equal to 0.25 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cell...
Specifications
Specifications
| Antigen | Themis |
| Applications | Flow Cytometry |
| Classification | Monoclonal |
| Clone | 1TMYS |
| Concentration | 0.2 mg/mL |
| Conjugate | eFluor 660 |
| Formulation | PBS with 0.09% sodium azide; pH 7.2 |
| Gene | Themis |
| Gene Accession No. | Q8BGW0 |
| Gene Alias | bA325O24.3; bA325O24.4; C6orf190; C6orf207; E430004N04Rik; FLJ40584; Gasp; GRB2-associated protein; Grb2-associating protein; MGC163388; Protein THEMIS; Protein thylex; RGD1560849; signaling phosphoprotein specific for T cells; SPOT; THEMIS; thylex; thymocyte expressed molecule involved in selection; thymocyte selection associated; thymocyte selection pathway associated; thymocyte-expressed molecule involved in selection; Tsepa |
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Frequently Asked Questions (FAQs)
Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).
Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.
The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.
The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.
As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.
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