Shop All Secondary Detection Kits & Reagents

Pluripotent Stem Cell Immunocytochemistry Kit (OCT4, SSEA4) (Invitrogen™)

The Pluripotent Stem Cell Immunocytochemistry Kit (OCT4, SSEA4) enables optimal image-based analysis of two key markers of human pluripotent stem cells (PSCs): OCT4 and SSEA4. This high performance immunocytochemistry (ICC) kit includes a complete set of primary and secondary antibodies, a nuclear DNA stain, and premade buffers for an optimized staining experiment. To characterize PSCs with other marker combinations, we recommend the use of the Pluripotent Stem Cell 4-Marker Immunocytochemistry Kit or the Pluripotent Stem Cell Immunocytochemistry Kit (SOX2, TRA-1-60).

With the Pluripotent Stem Cell Immunocytochemistry Kit (OCT4, SSEA4), you can:
• Confirm expression of key pluripotent stem cell markers with highly specific antibodies
• Get more information per sample by measuring more than one marker at a time
• Generate beautiful multiplexed images with minimal effort
• Save time and avoid cell loss by minimizing unnecessary wash steps in your staining protocol
• Process samples with confidence using a complete and optimized set of ICC reagents

Confirm Expression of Key PSC Markers
The antibodies included in the kit have been carefully selected and validated for high performance in the ICC analysis of PSCs, whether cultured under feeder-dependent or feeder-free media systems. OCT4 is a critical nuclear marker, while SSEA4 is an important cell surface marker.

Get More Information Per Sample
The antibodies in the kit have been designed to allow for the specific and simultaneous assessment of OCT4 and SSEA4 along with nuclear DNA staining to help save time and precious sample.

Generate Beautiful Multiplexed Images
Powered by the superior performance of bright, photostable Alexa Fluor® dyes, this kit enables the acquisition of beautiful multiplexed images of PSC marker expression using commonly available blue, green, and orange/red filter sets.

Minimize Unnecessary Wash Steps
The streamlined method recommended for this kit eliminates unnecessary wash steps found in standard ICC protocols to help save time and decrease the likelihood of cell loss during the staining protocol, especially when using glass coverslips or chamber slides. The only washes required are after incubation with primary and secondary antibodies.

Process Samples with Confidence
An optimized set of premade fixation, permeabilization, blocking, and wash buffers allows you to focus on answering scientific questions rather than spending time preparing reagents and determining compatibility.

What You Get
Easily scaled to any culture plate, dish, or slide format, the Pluripotent Stem Cell Immunocytochemistry Kit (OCT4, SSEA4) comes with reagents sufficient to stain 40 samples using a 50 uL staining volume. (See kit content details below.)

Alexa Fluor™ 647 Tyramide SuperBoost™ Kit, goat anti-rabbit IgG (Invitrogen™)

SuperBoost™ tyramide signal amplification is the most sensitive method for detection of low abundant targets in multiplexable fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC ), and in situ hybridization (ISH). SuperBoost kits combine the brightness of AlexaFluor™ dyes with the superior signal amplification of a poly-HRP-mediated tyramide labeling reaction to produce a sensitivity 10-200 times greater than standard methods. SuperBoost kit sensitivity is also 2-10 times greater than regular tyramide amplification techniques like TSA™. For standout research, SuperBoost kits sharpen your results for clear visibility into critical areas that standard imaging methods fail to reveal.

SuperBoost kits are simple to use and easily adapted to standard ICC, IHC, or FISH experimental protocols, using any cell or tissue type. Cells labeled using a SuperBoost kit can be imaged using any type of microscope, producing high-resolution multiplex images. This particular kit features AlexaFluor 647 tyramide (650/688 ex/em), detected using a standard Deep Red/Cy5 filter cube. This kit also features poly-HRP-conjugated goat anti-rabbit IgG secondary antibody.

Features of the SuperBoost kits include:
• Superior sensitivity for detection of low-level or hard-to-detect targets by fluorescent imaging
• Simple protocol and detection using standard filters
• Suitable for high-resolution multiplex images—co-label with DAPI, secondary antibodies, and other SuperBoost kits
• Requires 10-100 times less primary antibody then 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 using a SuperBoost kit requires 10-100 times less primary antibody then standard ICC/IHC/ISH experiments. SuperBoost kits offer superior specific signal intensity over background, so the protocol is easily optimized to detect specific signal in samples where high endogenous autofluorescence is observed.

Benefits of SuperBoost kits

Enhancement of signal using Alexa Fluor tyramides: SuperBoost kits utilize Alexa Fluor tyramides, which react with HRP to ultimately deposit bright and photostable Alexa Fluor dye on surrounding proteins and other similar molecules. SuperBoost kits are the only kits that combine the brightness of Alexa Fluor dyes with the enhancement of tyramide signal amplification to produce a superior signal.

Poly-HRP enhancement: Unlike TSA, SuperBoost kits employ poly-HRP-conjugated secondary antibodies. In such systems, several HRP enzymes are conjugated with short polymers, enhancing the signal by several fold over regular HRP systems. The poly-HRP is structured in such a way that the antibodies penetrate cells or tissue as efficiently as regular HRP-conjugated secondary antibodies. The molar enzyme/antibody protein ratio has an average 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 halt the HRP reaction. HRP stop solution can be used to obtain maximum signal, without increase of 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.

Reduction of background: SuperBoost kits include blockers for the elimination or reduction of endogenous peroxidase and fluorescent background signals. These blockers help ensure that only specific signals are enhanced while keeping non-specific/background signals in check.

CD44 Alexa Fluor™ 488 Conjugate Kit for Live Cell Imaging (Invitrogen™)

Molecular Probes® human pluripotent stem cell (PSC) live imaging kits provide reagents for superior imaging of live human PSCs. This CD44 Alexa Fluor® 488 Conjugate Kit includes an optimally labeled primary antibody paired with reduced-background imaging medium (FluoroBrite™ DMEM) for the specific detection of the negative PSC marker CD44. For fixed cell staining of PSC markers and germ layer markers, we recommend the Pluripotent Stem Cell 4-Marker Immunocytochemistry Kit and the 3-Germ Layer Immunocytochemistry Kit.

With the CD44 Alexa Fluor® 488 Conjugate Kit for Live Cell Imaging, you can:
• Detect the expression of a negative pluripotent stem cell marker using an optimally dye-conjugated antibody
• Preserve cell health and maximize fluorescence signal with FluoroBrite™ DMEM imaging medium
• Stain live cell samples with confidence using reagents that are tested to be free of common contaminants
• Combine with additional markers to get more information per sample

Detect Expression of a Negative PSC Marker
Powered by the high performance of bright, photostable Alexa Fluor® dyes, this kit enables reliable live-cell detection of CD44. CD44 is absent in PSCs and expressed on the surface of many other cell types, including human and mouse fibroblasts. This antibody conjugate has been demonstrated to show superior staining results when used with FluoroBrite™ DMEM.

Preserve Cell Health and Maximize Signal
Staining cells in FluoroBrite™ DMEM maximizes cell health and enhances signal-to-noise ratio when performing live-cell fluorescence imaging.

Stain Live-cell Samples with Confidence
Unlike most other conjugated antibodies, the reagents provided in this kit are 0.2 µm sterile-filtered and tested to be free of common contaminants.

Combine with Other Markers
To get more information per sample, combine this kit with the TRA-1-60 Alexa Fluor® 555 Conjugate Kit for Live Cell Imaging or the TRA-1-60 Alexa Fluor® 594 Conjugate Kit for Live Cell Imaging.

What You Get
Easily scaled to any culture plate, dish, or slide format, the CD44 Alexa Fluor® 488 Conjugate Kit comes with reagents sufficient to stain 50 samples using a 200 µL staining volume and includes:
• 200 µL of Alexa Fluor® 488 Rat Anti-Human and Mouse CD44 (50X concentration)
• 500 mL FluoroBrite™ DMEM

FMAT Blue™ Monofunctional Reactive Dye Labeling Kit (Applied Biosystems™)

The FMAT Blue® Monofunctional Dye Labeling Kit is supplied as a NHS-ester. The dye is a channel 1 dye emitting in the 650 nm range. The dye is designed to label antibodies, proteins and other biopolymers that have a primary amine, for use as secondary detection reagents.

For Research Use Only. Not for use in diagnostics procedures.

TRA-1-60 Alexa Fluor™ 488 Conjugate Kit for Live Cell Imaging (Invitrogen™)

Molecular Probes® human pluripotent stem cell (PSC) live imaging kits provide reagents for superior imaging of live human PSCs. This TRA-1-60 Alexa Fluor® 488 Conjugate Kit includes an optimally labeled primary antibody paired with reduced-background imaging medium (FluoroBrite™ DMEM) for the specific detection of the PSC marker TRA-1-60. For fixed cell staining of PSC markers and germ layer markers, we recommend the Pluripotent Stem Cell 4-Marker Immunocytochemistry Kit.

With the TRA-1-60 Alexa Fluor® 488 Conjugate Kit for Live Cell Imaging, you can:
• Confirm the expression of a key pluripotent stem cell marker using an optimally dye-conjugated antibody
• Preserve cell health and maximize fluorescence signal with FluoroBrite™ DMEM
• Stain live cell samples with confidence using reagents that are tested to be free of common contaminants

Confirm Expression of a Key PSC Marker
Powered by the high performance of bright, photostable Alexa Fluor® dyes, this kit enables reliable live-cell detection of the surface PSC marker TRA-1-60. This antibody conjugate has been demonstrated to show superior staining results when used with FluoroBrite™ DMEM.

Preserve Cell Health and Maximize Signal
Staining cells in FluoroBrite™ DMEM maximizes cell health and enhances signal-to-noise ratio when performing live-cell fluorescence imaging.

Stain Live-cell Samples with Confidence
Unlike most other conjugated antibodies, the reagents provided in this kit are 0.2 µm sterile-filtered and tested to be free of common contaminants.

What You Get
Easily scalable to any culture plate, dish, or slide format, the TRA-1-60 Alexa Fluor® 488 Conjugate Kit comes with reagents sufficient to stain 50 samples using a 200 µL staining volume and includes:
• 200 µL of Alexa Fluor® 488 Mouse Anti-Human TRA-1-60 (50X concentration)
• 500 mL FluoroBrite™ DMEM

Biotin XX Tyramide SuperBoost™ Kit, Streptavidin (Invitrogen™)

SuperBoost™ tyramide signal amplification is the most sensitive method for detection of low abundant targets in multiplexable fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC ), and in situ hybridization (ISH). SuperBoost kits combine the brightness of AlexaFluor™ dyes with the superior signal amplification of a poly-HRP-mediated tyramide labeling reaction to produce a sensitivity 10-200 times greater than standard methods. For standout research, SuperBoost kits sharpen your results for clear visibility into critical areas that standard imaging methods fail to reveal.

SuperBoost kits are simple to use and easily adapted to standard ICC, IHC, or FISH experimental protocols, using any cell or tissue type. Cells labeled using a SuperBoost kit can be imaged using any type of microscope, producing high-resolution multiplex images. This particular kit features biotin XX tyramide with HRP-conjugated streptavidin. For detection and additional signal amplification, conjugated streptavidin is used to detect specifically deposited biotin XX .

Features of the SuperBoost kits include:
• Superior sensitivity for detection of low-level or hard-to-detect targets by fluorescent imaging
• Simple protocol and detection using standard filters
• Suitable for high-resolution multiplex images—co-label with DAPI, secondary antibodies, and other SuperBoost kits
• Requires 10-100 times less primary antibody then 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 using a SuperBoost kit requires 10-100 times less primary antibody then standard ICC/IHC/ISH experiments. SuperBoost kits offer superior specific signal intensity over background, so the protocol is easily optimized to detect specific signal in samples where high endogenous autofluorescence is observed.

3-Germ Layer Immunocytochemistry Kit (Invitrogen™)

The 3-Germ Layer Immunocytochemistry Kit enables optimal image-based analysis of spontaneously differentiated embryoid bodies derived from human pluripotent stem cells. It detects widely accepted markers characteristic of the three embryonic germ layers: beta-III tubulin (TUJ1) for ectoderm, smooth muscle actin (SMA) for mesoderm, and alpha-fetoprotein (AFP) for endoderm. This high performance immunocytochemistry (ICC) kit includes a complete set of primary and secondary antibodies, a nuclear DNA stain, and premade buffers for an optimized staining experiment. To characterize undifferentiated pluripotent stem cells, we recommend use of the Pluripotent Stem Cell 4-Marker Immunocytochemistry Kit.

With the 3-Germ Layer Immunocytochemistry Kit, you can:
• Confirm expression of common germ layer markers with highly specific antibodies
• Get more information per sample by measuring more than one marker at a time
• Generate beautiful multiplexed images with minimal effort
• Save time and avoid cell loss by minimizing unnecessary wash steps in your staining protocol
• Process samples with confidence using a complete and optimized set of ICC reagents

Confirm Expression of Common Germ Layer Markers
The marker antibodies included in the kit have been carefully selected and validated for high performance in the ICC analysis of the three germ lineages in randomly differentiated embryoid bodies. TUJ1, SMA and AFP are markers that are widely used for the detection of neurons (ectoderm), cardiomyocytes (mesoderm), and primitive hepatocytes (endoderm), respectively.

Get More Information Per Sample
To help save time and precious sample, the antibody combinations in the kit have been designed to allow for the specific and simultaneous assessment of two to three markers along with nuclear DNA staining, depending on the usage of the far red filter.

Generate Beautiful Multiplexed Images
Powered by the superior performance of bright, photostable Alexa Fluor® dyes, this kit enables the acquisition of beautiful multiplexed images using commonly available blue, green, orange/red, and far red filter sets.

Minimize Unnecessary Wash Steps
The streamlined method recommended for this kit eliminates unnecessary wash steps found in standard ICC protocols to help save time and decrease the likelihood of cell loss during the staining protocol, especially when using glass coverslips or chamber slides. The only washes required are after incubation with primary and secondary antibodies.

Process Samples with Confidence
An optimized set of premade fixation, permeabilization, blocking, and wash buffers allows you to focus on answering scientific questions rather than spending time preparing reagents and determining compatibility.

What You Get
Easily scaled to any culture plate, dish, or slide format, the 3-Germ Layer Immunocytochemistry Kit comes with reagents sufficient to stain 20 samples using a 200 µL staining volume. (See kit content details below.)

LI Silver (LIS) Enhancement Kit (Invitrogen™)

The LI Silver (LIS) Enhancement Kit is a convenient, light-insensitive silver enhancement system for use with our colloidal gold and NANOGOLD reagents. Gold particles in the presence of silver (I) ions and a reducing agent act as catalysts to reduce the silver (I) ions to metallic silver. The metallic silver is deposited onto the gold, enlarging the particles to between 30 and 100 nm in diameter. Tissue or blots stained with colloidal gold are "developed" by this autometallographic procedure to give black staining that can be seen in the light microscope. The LI Silver Enhancement Kit is prepared for Molecular Probes by Nanoprobes, Inc.

Alexa Fluor™ 594 Tyramide SuperBoost™ Kit, goat anti-rabbit IgG (Invitrogen™)

SuperBoost™ tyramide signal amplification is the most sensitive method for detection of low abundant targets in multiplexable fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC ), and in situ hybridization (ISH). SuperBoost kits combine the brightness of AlexaFluor™ dyes with the superior signal amplification of a poly-HRP-mediated tyramide labeling reaction to produce a sensitivity 10-200 times greater than standard methods. SuperBoost kit sensitivity is also 2-10 times greater than regular tyramide amplification techniques like TSA™. For standout research, SuperBoost kits sharpen your results for clear visibility into critical areas that standard imaging methods fail to reveal.

SuperBoost kits are simple to use and easily adapted to standard ICC, IHC, or FISH experimental protocols, using any cell or tissue type. Cells labeled using a SuperBoost kit can be imaged using any type of microscope, producing high-resolution multiplex images. This particular kit features AlexaFluor 594 tyramide (591/617 ex/em), detected using a standard Red/Texas Red™ filter cube. This kit also features poly-HRP-conjugated goat anti-rabbit IgG secondary antibody.

Features of the SuperBoost kits include:
• Superior sensitivity for detection of low-level or hard-to-detect targets by fluorescent imaging
• Simple protocol and detection using standard filters
• Suitable for high-resolution multiplex images—co-label with DAPI, secondary antibodies, and other SuperBoost kits
• Requires 10-100 times less primary antibody then 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 using a SuperBoost kit requires 10-100 times less primary antibody then standard ICC/IHC/ISH experiments. SuperBoost kits offer superior specific signal intensity over background, so the protocol is easily optimized to detect specific signal in samples where high endogenous autofluorescence is observed.

Benefits of SuperBoost kits

Enhancement of signal using Alexa Fluor tyramides: SuperBoost kits utilize Alexa Fluor tyramides, which react with HRP to ultimately deposit bright and photostable Alexa Fluor dye on surrounding proteins and other similar molecules. SuperBoost kits are the only kits that combine the brightness of Alexa Fluor dyes with the enhancement of tyramide signal amplification to produce a superior signal.

Poly-HRP enhancement: Unlike TSA, SuperBoost kits employ poly-HRP-conjugated secondary antibodies. In such systems, several HRP enzymes are conjugated with short polymers, enhancing the signal by several fold over regular HRP systems. The poly-HRP is structured in such a way that the antibodies penetrate cells or tissue as efficiently as regular HRP-conjugated secondary antibodies. The molar enzyme/antibody protein ratio has an average 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 halt the HRP reaction. HRP stop solution can be used to obtain maximum signal, without increase of 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.

Reduction of background: SuperBoost kits include blockers for the elimination or reduction of endogenous peroxidase and fluorescent background signals. These blockers help ensure that only specific signals are enhanced while keeping non-specific/background signals in check.

Alexa Fluor™ 594 Goat Anti-Mouse SFX Kit, highly cross-adsorbed (Invitrogen™)

Alexa Fluor® SFX Kits contain Image-iT® FX signal enhancer (Cat. no. I36933) plus one of sixteen different Alexa Fluor® dye-labeled secondary antibodies. These kits provide 400 µg (0.2 mL of 2 mg/mL) of either goat anti-mouse IgG or goat anti-rabbit IgG antibody (as a standard or highly cross-adsorbed preparation) conjugated to Alexa Fluor® 488, Alexa Fluor® 555, Alexa Fluor® 594, or Alexa Fluor® 647 dye, four of our most commonly used Alexa Fluor® dyes. The Alexa Fluor® 594 goat anti-mouse IgG, highly cross-adsorbed, included with this kit is also available in a 1000 µg unit size (Cat. no. A11032).

Pierce™ Alkaline Phosphatase (Thermo Scientific™)

This purified calf intestinal alkaline phosphatase (CIP) is supplied in Tris buffer and 50% glycerol for use in protein research methods. The main applications for alkaline phosphatase in molecular biology and protein research are to remove 5'-phosphate groups from DNA or as a reporter system for immunoassays such as ELISA. For these latter methods, the enzyme is usually conjugated to specific primary or secondary antibodies and its activity is detected with a color-forming (or light-generating) substrate.

Features of Alkaline Phosphatase:

Purified form—ready to dilute and conjugate without prior dialysis
Concentrated—approximately 20 mg/mL (lot-specific value reported)
High specific activity—typically greater than 1600 units/mg (lot-specific value reported)
Tris buffered solution—supplied in 5 mM Tris, 5 mM magnesium chloride and 0.1 mM zinc chloride,
pH ~7.0 in 50% glycerol

One unit equals the amount of protein required to hydrolyze one micromole of p-nitrophenyl phosphate (PNPP) per minute at 25°C in a glycine buffer, pH 9.6.

Alexa Fluor™ 488 Tyramide SuperBoost™ Kit, streptavidin (Invitrogen™)

SuperBoost™ tyramide signal amplification is the most sensitive method for detection of low abundant targets in multiplexable fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC ), and in situ hybridization (ISH). SuperBoost kits combine the brightness of AlexaFluor™ dyes with the superior signal amplification of a poly-HRP-mediated tyramide labeling reaction to produce a sensitivity 10-200 times greater than standard methods. For standout research, SuperBoost kits sharpen your results for clear visibility into critical areas that standard imaging methods fail to reveal.

SuperBoost kits are simple to use and easily adapted to standard ICC, IHC, or FISH experimental protocols, using any cell or tissue type. Cells labeled using a SuperBoost kit can be imaged using any type of microscope, producing high-resolution multiplex images. This particular kit features AlexaFluor 488 tyramide (496/524 ex/em), detected using a standard Green/FITC/GFP filter cube. This kit also features HRP-conjugated streptavidin.

Features of the SuperBoost kits include:
• Superior sensitivity for detection of low-level or hard-to-detect targets by fluorescent imaging
• Simple protocol and detection using standard filters
• Suitable for high-resolution multiplex images—co-label with DAPI, secondary antibodies, and other SuperBoost kits
• Requires 10-100 times less primary antibody then 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 using a SuperBoost kit requires 10-100 times less primary antibody then standard ICC/IHC/ISH experiments. SuperBoost kits offer superior specific signal intensity over background, so the protocol is easily optimized to detect specific signal in samples where high endogenous autofluorescence is observed.

Benefits of SuperBoost kits

Enhancement of signal using Alexa Fluor tyramides: SuperBoost kits utilize Alexa Fluor tyramides, which react with HRP to ultimately deposit bright and photostable Alexa Fluor dye on surrounding proteins and other similar molecules. SuperBoost kits are the only kits that combine the brightness of Alexa Fluor dyes with the enhancement of tyramide signal amplification to produce a superior signal.

Reaction stop solution: Like any enzyme-based labeling system, it is possible to overdevelop the signal. SuperBoost kits include an HRP stop solution to halt the HRP reaction. HRP stop solution can be used to obtain maximum signal, without increase of 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.

Reduction of background: SuperBoost kits include blockers for the elimination or reduction of endogenous peroxidase and fluorescent background signals. These blockers help ensure that only specific signals are enhanced while keeping non-specific/background signals in check.

TSA™ Kit #1, with HRP—Goat Anti-Mouse IgG and biotin-XX Tyramide (Invitrogen™)

New SuperBoost™ Tyramide signal Amplification Kits offer 2-10 times the sensitivity of TSA™ kits. Explore the advantages of Biotin XX Tyramide SuperBoost™ Kit - Goat anti-Mouse IgG, which is replacing TSA™ Kit #1, with HRP--Goat Anti-Mouse IgG and biotin-XX Tyramide.

Tyramide signal amplification is an enzyme-mediated detection method that uses the catalytic activity of horseradish peroxidase (HRP) to generate high-density labeling of a target protein or nucleic acid sequence in situ. The TSA Kit #1 includes a HRP—goat anti-mouse IgG and biotin-XX tyramide. The deposited biotin molecules can be detected with any of our avidin, streptavidin or neutravidin conjugates.

Biotin-XX Tyramide Reagent (Invitrogen™)

Biotin XX Tyramide Reagent is a component of SuperBoost™ tyramide signal amplification kits made available separately here. This reagent can be used with any antibody conjugated to HRP for tyramide signal amplification for detection of low abundance targets in multiplexable fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC ), and in situ hybridization (ISH). For detection and additional signal amplification, conjugated streptavidin should be used to detect specifically deposited biotin XX.

SuperBoost reagents, when used as directed, can increase sensitivity up to 200 times over standard imaging methods. For standout research, SuperBoost reagents sharpen your results for clear visibility into critical areas that standard imaging methods fail to reveal. SuperBoost reagents are simple to use and easily adapted to standard ICC, IHC, or FISH experimental protocols, using any cell or tissue type. Cells labeled using a SuperBoost reagent can be imaged using any type of microscope, producing high-resolution multiplex images.

Features of the SuperBoost reagents include:
• Superior sensitivity for detection of low-level or hard-to-detect targets by fluorescent imaging
• Simple protocol and detection using standard filters
• Suitable for high-resolution multiplex images—co-label with TO-PRO™-3, secondary antibodies, and other SuperBoost kits
• Requires 10-100 times less primary antibody then standard ICC/IHC/ISH experiments

Alexa Fluor™ 647 Tyramide SuperBoost™ Kit, goat anti-mouse IgG (Invitrogen™)

SuperBoost™ tyramide signal amplification is the most sensitive method for detection of low abundant targets in multiplexable fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC ), and in situ hybridization (ISH). SuperBoost kits combine the brightness of AlexaFluor™ dyes with the superior signal amplification of a poly-HRP-mediated tyramide labeling reaction to produce a sensitivity 10-200 times greater than standard methods. SuperBoost kit sensitivity is also 2-10 times greater than regular tyramide amplification techniques like TSA™. For standout research, SuperBoost kits sharpen your results for clear visibility into critical areas that standard imaging methods fail to reveal.

SuperBoost kits are simple to use and easily adapted to standard ICC, IHC, or FISH experimental protocols, using any cell or tissue type. Cells labeled using a SuperBoost kit can be imaged using any type of microscope, producing high-resolution multiplex images. This particular kit features AlexaFluor 647 tyramide (650/688 ex/em), detected using a standard Deep Red/Cy5 filter cube. This kit also features poly-HRP-conjugated goat anti-mouse IgG secondary antibody.

Features of the SuperBoost kits include:
• Superior sensitivity for detection of low-level or hard-to-detect targets by fluorescent imaging
• Simple protocol and detection using standard filters
• Suitable for high-resolution multiplex images—co-label with DAPI, secondary antibodies, and other SuperBoost kits
• Requires 10-100 times less primary antibody then 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 using a SuperBoost kit requires 10-100 times less primary antibody then standard ICC/IHC/ISH experiments. SuperBoost kits offer superior specific signal intensity over background, so the protocol is easily optimized to detect specific signal in samples where high endogenous autofluorescence is observed.

Benefits of SuperBoost kits

Enhancement of signal using Alexa Fluor tyramides: SuperBoost kits utilize Alexa Fluor tyramides, which react with HRP to ultimately deposit bright and photostable Alexa Fluor dye on surrounding proteins and other similar molecules. SuperBoost kits are the only kits that combine the brightness of Alexa Fluor dyes with the enhancement of tyramide signal amplification to produce a superior signal.

Poly-HRP enhancement: Unlike TSA, SuperBoost kits employ poly-HRP-conjugated secondary antibodies. In such systems, several HRP enzymes are conjugated with short polymers, enhancing the signal by several fold over regular HRP systems. The poly-HRP is structured in such a way that the antibodies penetrate cells or tissue as efficiently as regular HRP-conjugated secondary antibodies. The molar enzyme/antibody protein ratio has an average 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 halt the HRP reaction. HRP stop solution can be used to obtain maximum signal, without increase of 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.

Reduction of background: SuperBoost kits include blockers for the elimination or reduction of endogenous peroxidase and fluorescent background signals. These blockers help ensure that only specific signals are enhanced while keeping non-specific/background signals in check.