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Flow cytometric analysis of TRIM expression on Human Peripheral Blood Mononuclear Cells (PBMC). Human PBMC were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with PE Mouse Anti-Human CD3 antibody (Cat. No. 555333) and with either Alexa Fluor™ 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Plot) or Alexa Fluor™ 647 Mouse Anti-TRIM antibody (Cat. No. 568558; Right Plot) at 5 µl/test. The bivariate pseudocolor density plot showing the correlated expression of TRIM versus CD3 (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Flow cytometric analysis of TRIM expression on Mouse splenic leucocytes. BALB/c mouse splenic leucocytes were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with PE Hamster Anti-Mouse CD3e antibody (Cat. No. 553063) and with either Alexa Fluor™ 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Plot) or Alexa Fluor™ 647 Mouse Anti-TRIM antibody (Cat. No. 568558) using an optimal dose of 1.25 µl/test. The bivariate pseudocolor density plot showing the correlated expression of TRIM versus CD3e (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ Alexa Fluor™ 647 Mouse Anti-TRIM
BD Pharmingen™ Alexa Fluor™ 647 Mouse Anti-TRIM
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
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配套商品
The TRIM-4 monoclonal antibody specifically recognizes TRIM which is also known as T cell receptor interacting molecule. TRIM is an ~30 kDa type III transmembrane protein that is expressed as a disulfide-linked homodimer. TRIM is comprised of a short extracellular domain (8 amino acids) followed by a transmembrane segment and cytosolic tail containing tyrosine motifs. These motifs may function as phosphorylation sites by Src- or Syk-family kinases after ligation of cell-surface receptors thus providing docking sites for SH2-domain-containing proteins. TRIM is encoded by TRAT1 (T cell receptor associated transmembrane adaptor 1) which belongs to the transmembrane adaptor protein (TRAP) family. TRIM is highly expressed by T cells with expression by some NK cells. TRIM functions as a signaling adaptor molecule that associates with the T-cell antigen receptor (TCR)/CD3 complex. TRIM appears to be involved in the regulation of TCR-mediated signaling. The TRIM-4 antibody reportedly crossreacts with mouse TRIM.
研发参考 (5)
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Huynh T, Würch A, Bruyns E, Korinek V, Schraven B, Eichmann K. Developmentally regulated expression of the transmembrane adaptor protein trim in fetal and adult T cells.. Scand J Immunol. 54(1-2):146-54. (Immunogen: Flow cytometry, Western blot). 查看参考
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Hübener C, Mincheva A, Lichter P, Schraven B, Bruyns E. Genomic organization and chromosomal localization of the human gene encoding the T-cell receptor-interacting molecule (TRIM).. Immunogenetics. 2000; 51(2):154-8. (Biology). 查看参考
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Kirchgessner H, Dietrich J, Scherer J, et al. The transmembrane adaptor protein TRIM regulates T cell receptor (TCR) expression and TCR-mediated signaling via an association with the TCR zeta chain.. J Exp Med. 2001; 193(11):1269-84. (Biology). 查看参考
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Leo A, Wienands J, Baier G, Horejsi V, Schraven B. Adapters in lymphocyte signaling.. J Clin Invest. 2002; 109(3):301-9. (Biology). 查看参考
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Tedoldi S, Paterson JC, Hansmann ML, et al. Transmembrane adaptor molecules: a new category of lymphoid-cell markers.. Blood. 2006; 107(1):213-21. (Biology). 查看参考
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