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ReadiUse™ Tyramide (TSA)/Styramide (PSA) Optimized Reaction Buffer

For many immunohistochemical (IHC) applications, the traditional enzymatic amplification procedures are sufficient for achieving adequate antigen detection. However, several factors limit the sensitivity and utility of these procedures. Tyramide signal amplification (TSA) has proven to be a particularly versatile and powerful enzyme amplification technique with improved assay sensitivity. TSA is based on the ability of HRP, in the presence of low concentrations of hydrogen peroxide, to convert labeled tyramine-containing substrate into an oxidized, highly reactive free radical that can covalently bind to tyrosine residues at or near the HRP. Power Styramide™ Signal Amplification (PSA™) is a recent upgrade to the TSA system with improved fluorescence signal 10-50 times higher than the corresponding TSA reagents. This HRP-induced radical reaction buffer is optimized for running both TSA and PSA reactions.

Example protocol

AT A GLANCE

Protocol summary
  1. Fix/permeabilize/block cells or tissue
  2. Add primary antibody in blocking buffer
  3. Add HRP-conjugated secondary antibody
  4. Prepare Styramide™ (PSA) or Tyramide (TSA) 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

Styramide™ (PSA) stock solution (100X)

Add 100 μL of DMSO into the vial of iFluor® dye-labeled Styramide™ conjugate (for example, Cat. 45000) to make 100X Styramide™ stock solution.

Note: Make single-use aliquots, and store unused 100X stock solution at 2-8 °C in a dark place and avoid repeat freeze-thaw cycles.

Hydrogen peroxide stock solution

Add 10 μL of 3% hydrogen peroxide (Component B) to 90 μL of ddH2O.

Note: Prepare the 100X H2O2 solution fresh on the day of use.

PREPARATION OF WORKING SOLUTION

Styramide™ (PSA) working solution (1X)

Every 1 mL of PSA working solution requires 1 mL TSA/PSA buffer (Component A), 10 μL of Styramide™ stock solution, and 10 μL of H2O2 stock solution.

Note: 1 mL is enough for 10 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.

Citations

View all 1 citations: Citation Explorer
Characterization of the Host Tissue Response Induced by a Biosynthetic Material Composed of Poly (4-Hydroxybutyrate)
Authors: Molina, Catalina Pineda
Journal: (2018)

References

View all 50 references: Citation Explorer
The autophagic response to Staphylococcus aureus provides an intracellular niche in neutrophils.
Authors: Prajsnar, Tomasz K and Serba, Justyna J and Dekker, Bernice M and Gibson, Josie F and Masud, Samrah and Fleming, Angeleen and Johnston, Simon A and Renshaw, Stephen A and Meijer, Annemarie H
Journal: Autophagy (2021): 888-902
Tagging the vanA gene in wastewater microbial communities for cell sorting and taxonomy of vanA carrying cells.
Authors: Gallego, Sara and Barkay, Tamar and Fahrenfeld, N L
Journal: The Science of the total environment (2020): 138865
Estimating the ploidy of Gracilariopsis lemaneiformis at both the cellular and genomic level1.
Authors: Chen, Haihong and Feng, Xiaoqing and Jiang, Minjie and Xiao, Baoheng and Zhang, Jingyu and Zhang, Wei and Hu, Yiyi and Sui, Zhenghong
Journal: Journal of phycology (2020): 1339-1348
A Protocol for Double Fluorescent In Situ Hybridization and Immunohistochemistry for the Study of Embryonic Brain Development in Tribolium castaneum.
Authors: Buescher, Marita and Oberhofer, Georg and Garcia-Perez, Natalia Carolina and Bucher, Gregor
Journal: Methods in molecular biology (Clifton, N.J.) (2020): 219-232
Detection of Cytokine Receptors Using Tyramide Signal Amplification for Immunofluorescence.
Authors: Wang, Herui and Pangilinan, Ryan L and Zhu, Yan
Journal: Methods in molecular biology (Clifton, N.J.) (2020): 89-97
Page updated on November 1, 2024

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Catalog Number45090
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Storage, safety and handling

Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12171501

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