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The antibody was purified by affinity chromatography and conjugated with PE under optimal conditions. The solution is free of unconjugated PE and unconjugated antibody.
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤0.5 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
COA:
Application Notes:
TW7-16B4 recognizes recombinant LAP, latent TGF-β, and pro-TGF-β. Additional reported applications (for relevant formats) include: Western blotting1 and immunoprecipitation1.
Application References:
1. Oida T, et al. 2010. PLoS ONE (FC, IP, WB) 2. Oida T, et al. 2011. PLoS ONE 6:e18365. (Neut)
C57BL/6 mouse splenocytes were stimulated with anti-mouse CD3, CD28, and recombinant mouse IL-2 for 48-hours, then surface stained with CD4 FITC and LAP (TGF-β1) (clone TW7-16B4) PE (top) or mouse IgG1, κ PE isotype control (bottom). This was followed by intracellular staining with FOXP3 Alexa Fluor® 647. Data shown was generated by gating on CD4+ lymphocyte population.
Transforming growth factor beta (TGF-β) is a cytokine that has critical functions in immune response by regulating Treg and Th17 cells. TGF-β is first synthesized as pro-TGF-β and then it is cleaved by furin proprotein convertase in the Golgi apparatus to produce the dimeric propeptides called latency-associated peptide (LAP) that non-covalently associates with the dimeric mature TGF-β to prevent its activity. This complex can further associate with latent-TGF-β-binding protein (LTBP) to produce a large latent form for deposition onto the extracellular matrix. The latent-TGF-β can be expressed on the membrane of activated Treg cells, immature dendritic cells, megakaryocytes, and platelets.
*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/terms). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.
This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
Purified anti-mouse LAP (TGF-β1)
C57BL/6 mouse splenocytes were stimulated with anti-mouse CD3, CD28, and recombinant mouse IL-2 for 48-hours, then surface stained with CD4 FITC and LAP (TGF-β1) (clone TW7-16B4) PE (top) or mouse IgG1, κ PE isotype control (bottom). This was followed by intracellular staining with FOXP3 Alexa Fluor® 647. Data shown was generated by gating on CD4+ lymphocyte population.
PE anti-mouse LAP (TGF-β1)
C57BL/6 mouse splenocytes were stimulated with anti-mouse CD3, CD28, and recombinant mouse IL-2 for 48-hours, then surface stained with CD4 FITC and LAP (TGF-β1) (clone TW7-16B4) PE (top) or mouse IgG1, κ PE isotype control (bottom). This was followed by intracellular staining with FOXP3 Alexa Fluor® 647. Data shown was generated by gating on CD4+ lymphocyte population.
APC anti-mouse LAP (TGF-β1)
CD3+CD28+IL-2-stimulated C57BL/6 mouse splenocytes (48 hours) were surface stained with CD4 FITC and LAP (clone TW7-16B4) APC (top) or mouse IgG1, κ APC isotype control (bottom), then intracellularly stained with FOXP3 PE. Data shown was generated by gating on CD4+ lymphocyte population.
