Shop All Secondary Detection Kits & Reagents

Pierce™ Horseradish Peroxidase (Thermo Scientific™)

Thermo Scientific Pierce Horseradish Peroxidase (HRP) is purified horseradish peroxidase enzyme for use in activity assays and conjugation to antibodies for ELISA, Western blot and immunohistochemistry applications.

Features of Thermo Scientific Pierce Horseradish Peroxidase:

Purified form—lyophilized salt-free powder; ready to dissolve and use
High specific activity—typically greater than 300 units/mg (lot-specific value reported)
Compared to AP—HRP is smaller (40kDa) than alkaline phosphatase (AP; 140kDa) and has higher specific enzyme activity than both AP and beta-galactosidase (b-Gal)
Many options—numerous substrate solutions and assay techniques are available for HRP

This purified horseradish peroxidase (HRP) is supplied lyophilized as a salt-free powder for reconstitution and use in protein research methods. The main application for HRP in molecular biology and protein research is as a reporter system for immunoassays and other probe-based assay techniques such as ELISA, Western blotting, EMSA and Southern blotting. The enzyme is usually conjugated to specific secondary antibodies or streptavidin, and its activity is detected with a color-forming (or light-generating) substrate.

One unit catalyses the production of 1 mg of purpurogallin from pyrogallol in 20 seconds at 20°C and pH 6.0.

Human Dopaminergic Neuron Immunocytochemistry Kit (Invitrogen™)

The Human Dopaminergic Neuron Immunocytochemistry Kit enables optimal image-based analysis of three key markers of neural differentiation: OTX2, FoxA2, and tyrosine hydroxylase (TH). 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.

The Human Dopaminergic Neuron Immunocytochemistry Kit enables you to:
• Confirm expression of key cellular 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 cellular markers
The antibodies included in the kit have been carefully selected and validated for high performance in the ICC analysis of floor plate progenitors and dopaminergic neurons. OTX2 and FoxA2 are critical nuclear markers while TH is an important cytoplasmic marker.

Get more information per sample
The antibody combinations in the kit have been designed to allow for the specific and simultaneous assessment of two markers at a time (e.g., OTX2 and FoxA2, and FoxA2 and TH) 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 key neural differentiation 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, helping to save time and decrease the likelihood of cell loss during the staining protocol. 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.

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.

TSA™ Kit #11, with HRP—Goat Anti-Rabbit 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-Rabbit IgG, which is replacing TSA™ Kit #11, with HRP--Goat Anti-Rabbit 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 #11 includes HRP—goat anti-rabbit IgG and biotin-XX tyramide The deposited biotin molecules can be detected with any of our avidin, streptavidin or neutravidin conjugates.

Alexa Fluor™ 555 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 555 tyramide (555/565 ex/em), detected using a standard Orange/RFP/TRITC 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.

SuperBoost™ Goat anti-Rabbit Poly HRP (Invitrogen™)

SuperBoost™ Goat anti-Rabbit Poly HRP is a component in SuperBoost™ tyramide signal amplification kits made available separately here. SuperBoost™ Goat anti-Rabbit Poly HRP can be used with any HRP sustrate such as tyramide, DAB for signal amplification and detection. The molar enzyme/antibody protein ratio has an average value of '4'. This reagent is provided at a ready-to-use 1X concentration, enough for 150 slides.

When used as directed, this reagent with other SuperBoost reagents can be used for detection of low abundant targets in multiplexable fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC ), and in situ hybridization (ISH) with up to 200 times more sensitivity than standard 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

Foxp3 Transcription Factor Staining Buffer Kit with Foxp3 Mouse Anti-Mouse mAb (clone 3G3), PE conjugate (Invitrogen™)

This product consists of a Foxp3 Transcription Factor Staining Buffer Kit (A24261) and a Foxp3 Mouse anti-Mouse mAb (clone 3G3), PE conjugate (A18690).

About the Antibody
The Foxp3 Mouse Anti-mouse mAb (clone 3G3), PE conjugate, is a monoclonal antibody that reacts with mouse Foxp3 protein. R-PE is a stable and highly soluble phycobiliprotein that provides maximal absorbance and fluorescence emission without susceptibility to internal or external fluorescence quenching, thus providing exceptional quantum yields and molar extinction coefficients. Use of R-PE in antibody conjugates results in probes with greatly enhanced detectability.

About the Buffers
The buffers in the included Foxp3 Transcription Factor Buffer Kit have been specially formulated to work with the PE conjugate of Foxp3 Mouse Anti-Mouse mAb (clone 3G3) for optimal observation of intranuclear staining when used in flow cytometry applications. The Foxp3 Transcription Factor Buffer Kit is compatible with intracellular cytokine staining within the same panel.

The kit contains 3 buffers:
• Foxp3 Fixation Transcription Factor and Permeabilization Buffer A (A24217, a 4X concentrate)
• Foxp3 Transcription Factor Fixation and Permeabilization Buffer B (A24218, a diluent)
• Foxp3 Transcription Factor Wash Buffer (A24261, 10X)

How the Buffers Work
The Foxp3 Transcription Factor Fixation and Permeabilization Buffer A is supplied as a 4X stock solution and must be diluted with Foxp3 Transcription Factor Fixation and Permeabilization Buffer B prior to use.

The Foxp3 Transcription Factor Wash Buffer is supplied as a 10X stock solution. The 10X stock solution should be diluted to a 1X working concentration in PBS containing 1% BSA and used for wash and intranuclear Foxp3 staining steps following fixation and permeabilization with 1X Foxp3 Fixation and Permabilization buffers.

About Foxp3
The 3G3 antibody reacts with mouse Foxp3 protein, a 50-55 kDa transcription factor that is a master regulator of regulatory T cell (Treg) development and function. Foxp3 is expressed constitutively by Tregs, which are further identified as being CD4+ CD25+, whereas resting conventional CD4+ T cells do not express Foxp3. However, TCR stimulation of naive T cells in the presence of TGF-beta has been shown to induce expression of Foxp3, driving their differentiation into Foxp3+ Tregs, which are called “induced" or “adaptive" Tregs. These cells are phenotypically similar to "natural" Tregs (CD4+ CD25+ Foxp3+), which originate in the thymus and comprise the majority of Tregs in the periphery. Tregs are critical for maintaining peripheral immune homeostasis. Defects in Foxp3 expression or Treg function have been implicated in the development of autoimmunity. Tregs have also been shown to restrict immune responses to infection, cancer, and vaccination. The 3G3 antibody may be used for intracellular detection of Foxp3 in cells from mouse and Rhesus macaque.

Related Links
Use the SpectraViewer to help you plan your experiments. Additional general information about Life Technologies® flow cytometry products and support is also available.

TSA™ Kit #2, with HRP—Goat Anti-Mouse IgG and Alexa Fluor™ 488 Tyramide (Invitrogen™)

New SuperBoost™ Tyramide signal Amplification Kits offer 2-10 times the sensitivity of TSA™ kits. Explore the advantages of Alexa Fluor™ 488 Tyramide SuperBoost™ Kit - Goat anti-Mouse IgG, which is replacing TSA™ Kit #2, with HRP--Goat Anti-Mouse IgG and Alexa Fluor® 488 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 #2 includes HRP—goat anti-mouse IgG and our exceptionally exceptionally bright, photostable, green-fluorescent Alexa Fluor 488 tyramide (which is spectrally similar to fluorescein).

TSA™ Kit #13, with HRP—Goat Anti-Rabbit IgG and Alexa Fluor™ 546 Tyramide (Invitrogen™)

New SuperBoost™ Tyramide signal Amplification Kits offer 2-10 times the sensitivity of TSA™ kits. Explore the advantages and superior replacement at Tyramide signal Amplification. 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 #13 includes HRP—goat anti-rabbit IgG and our bright, orange-fluorescent Alexa Fluor 546 tyramide (which is spectrally similar to Cy3 dye).

Alexa Fluor™ 594 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 594 tyramide (591/617 ex/em), detected using a standard Red/Texas Red™ 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.

Diaminobenzidine (DAB) Histochemistry Kit #3, with Streptavidin—HRP (Invitrogen™)

The DAB Histochemistry Kit for biotinylated probes contains sufficient materials to stain approximately 200 slides using horseradish peroxidase (HRP) and the HRP substrate DAB. The brown-colored DAB reaction product can be visualized by bright-fieldd light microscopy or, following osmication, by electron microscopy.

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.

Alexa Fluor™ 568 Signal-Amplification Kit for Mouse Antibodies (Invitrogen™)

Alexa Fluor® 568 Signal-Amplification Kit for Mouse Antibodies sensitively detects mouse primary antibodies. The kit contains an Alexa Fluor 568 rabbit anti—mouse IgG antibody conjugate, which is used to initially detect a mouse-derived primary antibody. It also contains an Alexa Fluor 568 goat anti—rabbit IgG antibody conjugate, which is used to dramatically amplify the fluorescence signal. Both conjugates have intense orange-red fluorescence with excitation/emission of ~578/603 nm, making them compatible with filters and settings appropriate for Lissamine rhodamine B or Rhodamine Red-X dye. The kit contains a detailed protocol and a sufficient amount of each reagent to stain 150—300 samples of adherent cells grown on coverslips. The kit also contains a protocol for use with flow cytometry.

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 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® 647 goat anti-mouse IgG, highly cross-adsorbed, included with this kit is also available in a 1000 µg unit size (Cat. no. A21236).