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.
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.
Product Details
BD Pharmingen™
TRAT1; TRIM; T cell receptor interacting molecule; TCRIM
Human (QC Testing), Mouse (Tested in Development)
Mouse IgG2a, κ
TRIM Cytoplasmic Sequence AA 29-186
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
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 the dye under optimum conditions and unreacted dye was removed.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- 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.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Transcription Factor Buffer Set RUO
Size
100 Tests
Cat No.
562574
Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control RUO
Size
50 µg
Cat No.
565365
PE Hamster Anti-Mouse CD3e RUO
Size
0.1 mg
Cat No.
553063
PE Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
555333
568558 Rev. 2
Antibody Details
TRIM-4
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.
568558 Rev. 2
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
568558 Rev.2
Citations & References
View product citations for antibody "568558" on CiteAb
Development References (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). View Reference
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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). View Reference
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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). View Reference
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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). View Reference
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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). View Reference
568558 Rev. 2
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.