iFluor® 750 Styramide *Superior Replacement for Alexa Fluor 750 tyramide*
Ordering information
Price | |
Catalog Number | |
Unit Size | |
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
Quotation | Request |
International | See distributors |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Molecular weight | 1257.67 |
Solvent | DMSO |
Spectral properties
Correction Factor (260 nm) | 0.044 |
Correction Factor (280 nm) | 0.039 |
Correction Factor (565 nm) | 0.0250 |
Correction Factor (650 nm) | 0.1413 |
Extinction coefficient (cm -1 M -1) | 2750001 |
Excitation (nm) | 757 |
Emission (nm) | 779 |
Quantum yield | 0.121 |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Freeze (< -15 °C); Minimize light exposure |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
See also: Antibodies and Proteomics, Antibody and Protein Labeling, Bioconjugation, Cell Structures and Organelles, Nucleus, DNA and RNA Quantitation, Fluorescent in situ hybridization (FISH), Horseradish Peroxidase (HRP) and Poly-HRP, iFluor® Dyes and Kits, Immunohistochemistry (IHC), Physiological Probes, Power Styramide™ Signal Amplification (PSA™)
Molecular weight 1257.67 | Correction Factor (260 nm) 0.044 | Correction Factor (280 nm) 0.039 | Correction Factor (565 nm) 0.0250 | Correction Factor (650 nm) 0.1413 | Extinction coefficient (cm -1 M -1) 2750001 | Excitation (nm) 757 | Emission (nm) 779 | Quantum yield 0.121 |
Power Styramide™ Signal Amplification (PSA™) system is one of the most sensitive methods that can detect extremely low-abundance targets in cells and tissues with improved fluorescence signal 10-50 times higher than the widely used tyramide (TSA) reagents. In combination with our superior iFluor® dyes that have higher florescence intensity, increased photostability and enhanced water solubility, the iFluor® dye-labeled Styramide™ conjugates can generate fluorescence signal with significantly higher precision and sensitivity (more than 100 times) than standard ICC/IF/IHC. PSA utilizes the catalytic activity of horseradish peroxidase (HRP) for covalent deposition of fluorophores in situ. PSA radicals have much higher reactivity than tyramide radicals, making the PSA system much faster, more robust and sensitive than the traditional TSA reagents. Compared to tyramide reagents, the Styramide™ conjugates have ability to label the target at higher efficiency and thus generate significantly higher fluorescence signal. Styramide™ conjugates also allow significantly less consumption of primary antibody compared to standard directly conjugate method or tyramide amplification with the same level of sensitivity. iFluor® 750 Styramide is a superior replacement for Alexa Fluor 750 tyramide or other spectrally similar fluorescent tyramide conjugates or TSA reagents.
Platform
Fluorescence microscope
Excitation | Cy7 filter set |
Emission | Cy7 filter set |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | Cy7 filter set |
Example protocol
AT A GLANCE
Protocol Summary
- Fix/permeabilize/block cells or tissue
- Add primary antibody in blocking buffer
- Add HRP-conjugated secondary antibody
- Prepare Styramide™ working solution and apply in cells or tissue for 5-10 minutes at room temperature
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
1. Styramide™ stock solution (100X)
Add 100 µL of DMSO into the vial of iFluor™ dye-labeled Styramide™ conjugate to make 100X Styramide™ stock solution. Note: Make single use aliquots, and store unused 100X stock solution at 2-8 °C in dark place and avoid repeat freeze-thaw cycles.2. H2O2 stock solution
Add 10 µL of 3% hydrogen peroxide (Not provided) to 90 µL of ddH2O. Note: Prepare the 100X H2O2 solution fresh on the day of use.PREPARATION OF WORKING SOLUTION
1. Styramide™ working solution (1X)
Every 1 mL of Reaction Buffer requires 10 µL of Styramide™ stock solution and 10 µL of H2O2 stock solution. Note: The Styramide™ provided is enough for 100 tests based on 100 µL of Styramide™ working solution needed per coverslip or per well in a 96-well microplate. Note: The Styramide™ working solution must be used within 2 hours after preparation and avoid direct exposure to light.2. Secondary antibody-HRP working solution
Make appropriate concentration of secondary antibody-HRP working solution as per the manufacturer's recommendations.SAMPLE EXPERIMENTAL PROTOCOL
This protocol is applicable for both cells and tissues staining.
Protocol can be found at https://www.aatbio.com/resources/guides/paraffin-embedded-tissueimmunohistochemistry-protocol.html
Cell fixation and permeabilization
- Fix the cells or tissue with 3.7% formaldehyde or paraformaldehyde, in PBS at room temperature for 20 minutes.
- Rinse the cells or tissue with PBS twice.
- Permeabilize the cells with 0.1% Triton X-100 solution for 1-5 minutes at room temperature.
- Rinse the cells or tissue with PBS twice.
Tissue fixation, deparaffinization and rehydration
Deparaffinize and dehydrate the tissue according to the standard IHC protocols. Perform antigen retrieval with preferred specific solution/protocol as needed.Protocol can be found at https://www.aatbio.com/resources/guides/paraffin-embedded-tissueimmunohistochemistry-protocol.html
Peroxidase labeling
- Optional: Quench endogenous peroxidase activity by incubating cell or tissue sample in peroxidase quenching solution (such as 3% hydrogen peroxide) for 10 minutes. Rinse with PBS twice at room temperature.
- Optional: If using HRP-conjugated streptavidin, it is advisable to block endogenous biotins by biotin blocking buffer.
- Block with preferred blocking solution (such as PBS with 1% BSA) for 30 minutes at 4 °C.
- Remove blocking solution and add primary antibody diluted in recommended antibody diluent for 60 minutes at room temperature or overnight at 4 °C.
- Wash with PBS three times for 5 minutes each.
- Apply 100 µL of secondary antibody-HRP working solution to each sample and incubate for 60 minutes at room temperature. Note: Incubation time and concentration can be varied depending on the signal intensity.
- Wash with PBS three times for 5 minutes each.
Styramide labeling
- Prepare and apply 100 µL of Styramide™ working solution to each sample and incubate for 5-10 minutes at room temperature. Note: If you observe non-specific signal, you can shorten the incubation time with Styramide. You should optimize the incubation period using positive and negative control samples at various incubation time points. Or you can use lower concentration of Styramide in the working solution.
- Rinse with PBS three times.
Counterstain and fluorescence imaging
- Counterstain the cell or tissue samples as needed. AAT provides a series of nucleus counterstain reagents as listed in Table 1. Follow the instruction provided with the reagents.
- Mount the coverslip using a mounting medium with anti-fading properties.
- Use the appropriate filter set to visualize the signal from the Styramide labeling.
Cat# | Product Name | Ex/Em (nm) |
17548 | Nuclear Blue™ DCS1 | 350/461 |
17550 | Nuclear Green™ DCS1 | 503/526 |
17551 | Nuclear Orange™ DCS1 | 528/576 |
17552 | Nuclear Red™ DCS1 | 642/660 |
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of iFluor® 750 Styramide *Superior Replacement for Alexa Fluor 750 tyramide* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 79.512 µL | 397.561 µL | 795.121 µL | 3.976 mL | 7.951 mL |
5 mM | 15.902 µL | 79.512 µL | 159.024 µL | 795.121 µL | 1.59 mL |
10 mM | 7.951 µL | 39.756 µL | 79.512 µL | 397.561 µL | 795.121 µL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum
Open in Advanced Spectrum Viewer


Spectral properties
Correction Factor (260 nm) | 0.044 |
Correction Factor (280 nm) | 0.039 |
Correction Factor (565 nm) | 0.0250 |
Correction Factor (650 nm) | 0.1413 |
Extinction coefficient (cm -1 M -1) | 2750001 |
Excitation (nm) | 757 |
Emission (nm) | 779 |
Quantum yield | 0.121 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
iFluor® 750 maleimide | 757 | 779 | 2750001 | 0.121 | 0.044 | 0.039 |
iFluor® 750 amine | 757 | 779 | 2750001 | 0.121 | 0.044 | 0.039 |
iFluor® 750 hydrazide | 757 | 779 | 2750001 | 0.121 | 0.044 | 0.039 |
iFluor® 350 Styramide *Superior Replacement for Alexa Fluor 350 tyramide* | 345 | 450 | 200001 | 0.951 | 0.83 | 0.23 |
iFluor® 488 Styramide *Superior Replacement for Alexa Fluor 488 tyramide and Opal 520* | 491 | 516 | 750001 | 0.91 | 0.21 | 0.11 |
iFluor® 546 Styramide *Superior Replacement for Alexa Fluor 546 tyramide* | 541 | 557 | 1000001 | 0.671 | 0.25 | 0.15 |
iFluor® 555 Styramide *Superior Replacement for Alexa Fluor 555 tyramide and Opal 570* | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
iFluor® 568 Styramide *Superior Replacement for Alexa Fluor 568 tyramide* | 568 | 587 | 1000001 | 0.571 | 0.34 | 0.15 |
iFluor® 594 Styramide *Superior Replacement for Alexa Fluor 594 tyramide* | 588 | 604 | 1800001 | 0.531 | 0.05 | 0.04 |
Show More (13) |
Images

Figure 1. Power Styramide™ Signal Amplification (PSA™) system is one of the most sensitive methods that can detect extremely low-abundance targets in cells and tissues with improved fluorescence signal 10-50 times higher than the widely used tyramide (TSA) reagents. In combination with our superior iFluor® dyes that have higher florescence intensity, increased photostability and enhanced water solubility, the iFluor® dye-labeled Styramide™ conjugates can generate fluorescence signal with significantly higher precision and sensitivity (more than 100 times) than standard ICC/IF/IHC. PSA utilizes the catalytic activity of horseradish peroxidase (HRP) for covalent deposition of fluorophores in situ. PSA radicals have much higher reactivity than tyramide radicals, making the PSA system much faster, more robust and sensitive than the traditional TSA reagents.

Figure 2. Fluorescence IHC of formaldehyde-fixed, paraffin-embedded human lung adenocarcinoma positive tissue using PSA ™ and TSA amplified methods. Human lung adenocarcinoma positive tissue sections were stained with Mouse anti-EpCAM or Mouse IgG antibody and then incubated with HRP-labeled Goat anti-Mouse IgG secondary antibody followed by iFluor® 750 Styramide™ (Cat#45065) or Alexa Fluor® 750 tyramide stain respectively. Images showed that iFluor® 750 PSA™ super signal amplification can increase the sensitivity of fluorescence IHC over Alexa Fluor® 750 TSA method.

Figure 3. Fluorescence images of HeLa cells labeled with Mouse anti-Tubulin primary antibody. Cells were then stained with a HRP-labeled Goat anti-Mouse IgG secondary antibody followed by iFluor® 750 Styramide™ (Left) or Alexa Fluor® 750 tyramide (Right), respectively. Fluorescence images were taken using the Cy7 filter set. iFluor® 750 Styramide™ shows significantly higher fluorescence intensity than Alexa Fluor® 750 tyramide if under the same exposure time.