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TrueBlack® WB Blocking Buffer Kit

The TrueBlack® WB Blocking Buffer Kit is a ready-to-use buffer system that provides optimal specificity, sensitivity, and background signal suppression for fluorescence-based western blotting (WB).

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50 membranes
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Product Description

The TrueBlack® WB Blocking Buffer Kit is a ready-to-use buffer system for fluorescence-based western blotting (WB). The buffers are designed to achieve optimal specificity and sensitivity by blocking background from the non-specific interaction between the fluorophore-labeled antibodies and the membrane.

  • Blocks non-specific protein binding, reducing background fluorescence over the entire membrane
  • Works as well or better than LI-COR’s Odyssey® Blocking Buffer, at a lower cost
  • Suppresses background caused by charged fluorescent dyes (performs better than BSA, gelatin, or casein)
  • Reduces antibody cross-reactivity, eliminating non-specific bands
  • Compatible with both polyvinylidene difluoride (PVDF) and nitrocellulose membranes
  • Can be used with antibodies labeled with all types of fluorophores spanning the visible and NIR spectrum
  • Contains non-mammalian blocking agents, for broad secondary antibody compatibility
  • TBS based buffer suitable for phosphoprotein detection
  • NOT designed for chemiluminescence detection using HRP conjugated antibodies

Non-specific signal in WB can arise from multiple sources, including antibody cross-reactivity with off-target proteins, non-specific antibody adsorption to the membrane, and membrane autofluorescence. Another potential cause of background that is not well-known is the effect of fluorescent dyes themselves on the specificity of labeled antibodies. Next-generation fluorescent dyes like Alexa Fluor® or CF® dyes often carry multiple negative charges to improve dye solubility and brightness of conjugates. However, the extra charge carried by the dye can result in non-specific antibody binding and background fluorescence. The TrueBlack® WB Blocking Buffer Kit blocks background from multiple sources including charged dye conjugates. It is especially advantageous for phosphoprotein detection, significantly improving specificity compared to conventional blocking buffers.

The TrueBlack® WB Blocking Buffer Kit belongs to our TrueBlack® line of background reducing agents for fluorescence applications, which includes TrueBlack® Lipofuscin Autofluorescence Quencher for tissue staining, and TrueBlack® IF Background Suppressor System (Permeabilizing) for blocking non-specific immunofluorescence staining.

When compared to LI-COR’s Odyssey® Blocking Buffer, the TrueBlack® WB Blocking Buffer works as well or better, and is priced lower on a per membrane basis. See below for a size comparison.

Comparison between TrueBlack® WB Blocking Buffer Kit and Odyssey® Blocking Buffer

ProductTrueBlack® WB Blocking
Buffer Kit
Odyssey® Blocking Buffer
Trial SizeFor 10 membranes125 mL for 4 membranes
Full SizeFor 50 membranes500 mL for 16 membranes
Number of membranes if Odyssey® Blocking Buffer is used for each blocking and antibody dilution step. TrueBlack® WB Blocking Buffer Kit includes enough buffers for all blocking and antibody incubation steps for the stated number of membranes.

 

Alexa Fluor is a registered trademark of Thermo Fisher Scientific. LI-COR and Odyssey are registered trademarks of LI-COR Inc.
 

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