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BV421 Mouse Anti-Mouse LAP
BV421 Mouse Anti-Mouse LAP
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 CD3e (Cat. No. 553057) and Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294) antibodies in the presence of Natural Transforming Growth Factor-b protein (TGF-b; Corning Cat. No. CB-40039). The cells were harvested and stained with PE Rat Anti-Mouse CD4 antibody (Cat. No. 553730/557308/561829) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-Mouse LAP antibody (Cat. No. 565638; 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 CD3e (Cat. No. 553057) and Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294) antibodies in the presence of Natural Transforming Growth Factor-b protein (TGF-b; Corning Cat. No. CB-40039). The cells were harvested and stained with PE Rat Anti-Mouse CD4 antibody (Cat. No. 553730/557308/561829) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-Mouse LAP antibody (Cat. No. 565638; 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
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BD Horizon™
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_2739315
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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  7. An isotype control should be used at the same concentration as the antibody of interest.
565638 Rev. 1
Antibody Details
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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.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

565638 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
565638 Rev.1
Citations & References
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Development References (5)

  1. 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: Flow cytometry, Immunoprecipitation, Western blot). View Reference
  2. 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
  3. 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
  4. 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
  5. 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
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565638 Rev. 1

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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.