Fluorescence background reducers: TrueBlack

The TrueBlack® reagents from Biotium offer rapid and simple blocking of fluorescent background from multiple sources for optimal specificity.

• TrueBlack IF Background Suppressor System blocks IF background from multiple sources
• TrueBlack Western Blot Blocking Buffer Kit offers superior blocking for fluorescent/near-IR west
• TrueBlack Lipofuscin Autofluorescence Quencher clears the way for immunofluorescence of human tissue
   

 Benefits of using TrueBlack IF background suppressor system
• Suppresses background from non-specific antibody binding and charged fluorescent dyes
• More efficient than Image-iT® FX, block & permeabilize in just 10 minutes
• Complete system for blocking, permeabilizing, and antibody dilution
• Non-mammalian blocking agents, for broad secondary antibody compatibility
• For immunofluorescence on cells or tissue sections

TrueBlack IF Background Suppressor reduces non-specific binding of antibodies conjugated to charged fluorescent dyes. Methanol-fixed HeLa cells were blocked for 10 minutes with fish gelatin blocking buffer or TrueBlack IF Background Suppressor, then stained with no primary antibody or mouse anti-tubulin antibody, followed by goat anti-mouse secondary antibody conjugated to the indicated dyes. Antibodies were diluted in the same buffer used for blocking

Benefits of using TrueBlack WB blocking buffer kit
• Blocks as well or better than Odyssey® Blocking Buffer, at a lower price
• Reduces non-specific protein bands and background over entire membrane
• Suppresses background from charged dyes better than BSA, gelatin, or casein
• Compatible with PVDF and nitrocellulose membranes
• Contains no mammalian proteins, for broad antibody compatibility
• For visible and near-IR fluorescent westerns

Western detection of phospho-Erk1/2 in PDGF-stimulated NIH-3T3 cell lysate. Membranes were blocked with fish gelatin blocking buffer, LI-COR® Odyssey® TBS Blocking Buffer, or TrueBlack WB Blocking Buffer. Phosphorylated Erk was detected using rabbit anti-pErk1/2 and CF®680R donkey anti-rabbit antibodies. TrueBlack WB Blocking Buffer gave lower background fluorescence and better specificity compared to the other buffers.

Benefits of using the TrueBlack Lipofuscin Autofluorescence Quencher
• Eliminates lipofuscin autofluorescence
• Reduces autofluorescence from non-lipofuscin sources
• Doesn’t cause high background
• Can be used before or after immunofluorescence staining
• Clears the way for fluorescence imaging of human and aged animal tissue

TrueBlack treatment before staining preserves antibody brightness. Human cerebral cortex was left untreated or treated with TrueBlack, then stained with CF®488A anti-NeuN (neuronal nuclei, green) and DAPI (nuclei, blue). In untreated tissue, lipofuscin granules appeared as white speckles (left). TrueBlack pre-treatment quenched lipofuscin fluorescence without affecting specific staining (right).

Material available for download
Fluorescence background blockers brochure
TrueBlack IF background suppressor system protocol
TrueBlack WB blocking buffer kit protocol
TrueBlack lipofuscin autofluorescence quencher protocol

TrueBlack citations

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