Overview
Amplification of the SuperBoost ™ tyramide signal is the most sensitive method for the detection of low abundant targets in multiplexable fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC), and in situ hybridization (ISH). SuperBoost kits combine the brilliance of AlexaFluor ™ dyes with the superior signal amplification of a poly-HRP-mediated tyramide labeling reaction to produce 10 to 200 times greater sensitivity than standard methods.
The sensitivity of the SuperBoost kit is also 2 to 10 times higher than common tyramide amplification techniques such as TSA ™. For outstanding research, SuperBoost kits enhance your results for clear visibility into critical areas that standard imaging methods fail to reveal.
SuperBoost kits are easy to use and easily adapt to standard ICC, IHC, or FISH experimental protocols, using any type of cell or tissue. Cells labeled with a SuperBoost kit can be obtained with any type of microscope, producing high-resolution multiplex images. This particular kit features Tyramide AlexaFluor 488 (496/524 ex/em), detected using a standard Green / FITC / GFP filter cube. This kit also includes poly-HRP conjugated goat anti-mouse IgG secondary antibody.
Features of the SuperBoost kits include:
- Superior sensitivity for detection of low-level or difficult-to-detect targets using fluorescent images
- Simple protocol and detection using standard filters
- Suitable for high-resolution multiplex imaging – co-tagged with DAPI, secondary antibodies, and other SuperBoost kits
- Requires 10 to 100 times fewer primary antibodies than standard ICC / IHC / ISH experiments
SuperBoost kits are based on the tyramide signal amplification system, which uses the catalytic activity of horseradish peroxidase (HRP) to generate high-density labeling of a target protein or nucleic acid sequence in situ.
A typical ICC / IHC / ISH experiment with a SuperBoost kit requires 10 to 100 times fewer primary antibodies than standard ICC / IHC / ISH experiments. SuperBoost kits offer superior specific signal intensity over the background, so the protocol is easily optimized to detect specific signals in samples where high endogenous autofluorescence is observed.
Benefits of SuperBoost Kits
Signal enhancement using Alexa Fluor tyramides –
SuperBoost kits use Alexa Fluor tyramides, which react with HRP to ultimately deposit the bright, photostable Alexa Fluor dye on surrounding proteins and other similar molecules. SuperBoost kits are the only kits that combine the brilliance of Alexa Fluor dyes with enhanced tyramide signal amplification to produce a superior signal.
Poly-HRP Enhancement –
Unlike TSA, SuperBoost kits employ poly-HRP-conjugated secondary antibodies. In such systems, various HRP enzymes are conjugated to short polymers, enhancing the signal several times more than regular HRP systems. Poly-HRP is structured in such a way that the antibodies penetrate cells or tissue with the same efficiency as regular HRP-conjugated secondary antibodies. The enzyme/antibody protein molar ratio has a mean value of “4”.
Reaction Stop Solution:
Like any enzyme-based labeling system, it is possible to overdevelop the signal. SuperBoost kits include an HRP stop solution to stop the HRP reaction. The HRP stop solution can be used to obtain the maximum signal, without increasing the background signal. Images produced with optimized HRP reaction times are as sharp as images produced with standard ICC / IHC / ISH methods, but with 10-200 times more sensitivity.
Background Reduction:
SuperBoost kits include blockers for endogenous peroxidase removal or reduction and fluorescent background signals. These jammers help ensure that only specific signals are enhanced while keeping non-specific / background signals in check.
