Two-color flow cytometric analysis of LAP expression on activated mouse CD4+ T cells. BALB/c mouse splenic leucocytes were activated in culture (2 days) with plate-bound Purified NA/LE Hamster Anti-Mouse CD3 (Cat. No. 553057) and Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294) antibodies in the presence of Purified Natural Transforming Growth Factor-b (TGF-b; Corning Cat. No. CB-40039). The cells were harvested and stained with BD Horizon™ BV421 Rat Anti-Mouse CD4 (Cat. No. 562891) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; Left Plot) or Alexa Fluor® 647 Mouse Anti-Mouse LAP antibody (Cat. No. 565636; Right Plot). Two-color flow cytometric dot plots show the correlated expression patterns of CD4 versus LAP (or Ig Isotype control staining) for gated events with the forward and side light-scatter characteristics of activated lymphocytes. Flow cytometry was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of LAP expression on activated mouse CD4+ T cells. BALB/c mouse splenic leucocytes were activated in culture (2 days) with plate-bound Purified NA/LE Hamster Anti-Mouse CD3 (Cat. No. 553057) and Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294) antibodies in the presence of Purified Natural Transforming Growth Factor-b (TGF-b; Corning Cat. No. CB-40039). The cells were harvested and stained with BD Horizon™ BV421 Rat Anti-Mouse CD4 (Cat. No. 562891) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; Left Plot) or Alexa Fluor® 647 Mouse Anti-Mouse LAP antibody (Cat. No. 565636; Right Plot). Two-color flow cytometric dot plots show the correlated expression patterns of CD4 versus LAP (or Ig Isotype control staining) for gated events with the forward and side light-scatter characteristics of activated lymphocytes. Flow cytometry was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Pharmingen™
Latency-associated peptide; TGFB1; TGFbeta; Transforming growth factor beta
Mouse (QC Testing)
Mouse IgG1, κ
Mouse Tgfb1-transduced Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2739314
Aqueous buffered solution containing ≤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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557714
Purified NA/LE Hamster Anti-Mouse CD3e RUO
Size
0.5 mg
Cat No.
553057
Purified NA/LE Hamster Anti-Mouse CD28 RUO
Size
0.5 mg
Cat No.
553294
BV421 Rat Anti-Mouse CD4 RUO
Size
50 µg
Cat No.
562891
565636 Rev. 1
Antibody Details
TW7-16B4
The TW7-16B4 monoclonal antibody specifically binds to Latency-Associated Peptide (LAP), a component of the dimeric Transforming Growth Factor-beta 1 (TGF-β1) propeptide encoded by Tgfb1. Prior to secretion, the dimeric LAP-TGF-β1 propeptide is cleaved resulting in a biologically inactive form of dimeric TGF-β1 that is noncovalently associated with dimeric LAP (latent TGF-β1). This complex may be expressed on the surface of TGF-β1-producing cells or be further processed by proteolytic removal of LAP to release the biologically active mature form of the soluble TGF-β1 homodimer. Platelets contain TGF-β1 and most nucleated cells, including tumor cells and cells that comprise the innate and adaptive immune system can produce TGF-β1. TGF-β1 is a potent multifunctional cytokine that regulates numerous processes including development, hematopoiesis, tissue remodeling, wound repair, and immunity as well as cancer and autoimmune diseases.
565636 Rev. 1
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 520-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
565636 Rev.1
Citations & References
Development References (5)
-
Oida T, Weiner HL. Overexpression of TGF-β1 gene induces cell surface localized glucose-regulated protein 78-associated latency-associated peptide/TGF-β. J Immunol. 2010; 185(6):3529-3535. (Clone-specific). View Reference
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Oida T, Weiner HL. TGF-β induces surface LAP expression on murine CD4 T cells independent of Foxp3 induction. PLoS ONE. 2010; 5(11):e15523. (Immunogen: Flow cytometry, Immunoprecipitation, Western blot). View Reference
-
Oida T, Zhang X, Goto M, et al. CD4+CD25- T cells that express latency-associated peptide on the surface suppress CD4+CD45RBhigh-induced colitis by a TGF-beta-dependent mechanism. J Immunol. 2003; 170(5):2516-2522. (Biology). View Reference
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Rubtsov YP, Rudensky AY. TGFbeta signalling in control of T-cell-mediated self-reactivity. Nat Rev Immunol. 2007; 7(6):443-453. (Biology). View Reference
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Wilkinson KA, Martin TD, Reba SM, et al. Latency-Associated Peptide of Transforming Growth Factor β enhances mycobacteriocidal immunity in the lung during Mycobacterium bovis BCG infection in C57BL/6 mice. Infect Immun. 2000; 68(11):6505-6508. (Biology). View Reference
565636 Rev. 1
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.
