Immunofluorescence (IF) is a powerful yet simple method for visualizing protein expression in tissues or cells using fluorophore-conjugated antibodies. Interfering background signal, such as autofluorescence, which is fluorescence inherent or endogenous to tissue, cell, or due to certain fixatives, can hamper your ability to accurately distinguish real target antigen expression.
Description
The TrueVIEW Autofluorescence Quenching Kit (also available with DAPI) provides a novel way to diminish unwanted autofluorescence from non-lipofuscin sources and dramatically improve signal-to-noise ratio. It removes unwanted autofluorescence in tissue sections due to aldehyde fixation, red blood cells, and structural elements such as collagen and elastin. The quenching action of the kit reagents provides a clear, unambiguous, “true view” localization of the target antigen.
Features:
- Effective on problematic tissues such as kidney and spleen
- Easy-to-use, one-step method
- Compatible with a wide selection of fluorophores
- Compatible with standard epifluorescence and confocal laser microscopes
- Availability with DAPI counterstain and without (Mounting medium contains DAPI for counterstaining in one step)
One kit is sufficient to treat approximately 100 to 150 tissue sections.
Available TrueVIEW Autofluorescence Quenching Kits:
- Vector TrueVIEW Autofluorescence Quenching Kit
- Vector TrueVIEW Autofluorescence Quenching Kit with DAPI
Tissue autofluorescence often occurs with aldehyde fixation or from inherent native tissue components (collagen, elastin, and red blood cells). The extent and intensity of autofluorescence background frequently makes it difficult or impossible to distinguish specific signals in immunofluorescence applications.
Most methods for reduction of tissue autofluorescence act primarily on lipofuscin granules, and are not broadly effective against the most common sources of autofluorescence targeted by TrueVIEW Quencher.
Current methods for reducing autofluorescence primarily include “home brew” concoctions such as sodium borohydride and other ink-based products. These methods are essentially ineffective against aldehyde induced autofluorescence. In contrast, Vector TrueVIEW reagent binds to hydrophilic compounds and effectively quenches endogenous autofluorescence.
Easy To Apply
Following completion of the IF staining procedure:
Mode of Action
Following completion of the IF staining procedure:
Comparison Between Vector TrueVIEW Reagent and Other Autofluorescence Reducing Agents
We compared the effectiveness of Vector TrueVIEW quenching action in parallel with other commercially available autofluorescence reducing products and “home brew” reagents, on serial sections of formalin-fixed, paraffin-embedded human pancreas visualized using a standard fluorescein (green) filter. No specific immunofluorescence staining was conducted. The images below highlight our results. All images were acquired under identical conditions (including microscope objective and exposure times).
Supplier
Vector Laboratories
Vector Laboratories is recognized as a pioneer and market leader in immunohistochemistry, immunofluorescence, glycobiology, and bioconjugation products.