Biotinyl tyramide 

Biotinyl tyramide is a biotin derivative used for tyramide signal amplification (TSA), as a reagent to amplify both immunohistochemical signals and in situ hybridization protocols. Biotinyl tyramide can be used for the research of tyramide signal amplification^{[1][2][3][4][5]}.

Guide (The following is our recommended protocol. This protocol is for reference only and should be modified according to your specific needs).
1. Solution preparation^[7]
1.1 Preparation of stock solution
Solvent: DMSO
Concentration: 27.5 mM (optimize according to the experiment).
Storage: Aliquot and store at -20°C or -80°C protected from light. Avoid repeated freeze-thaw cycles.
1.2 Preparation of working solution
Dilute with PBS or serum-free culture medium to 27.5 μM (optimize according to the experiment).
Note: The working solution should be freshly prepared, used immediately, and protected from light.

Biotinyl tyramide assay^[7].
1. Wash HaCaT cells and HaCaT DUOX1 knockout fusion cells on coverslips with culture medium. 2. Add 1 μM Thapsigargin (HY-13433), and incubate at 37°C for 5 min in the presence or absence of 0.2 U/mL horseradish peroxidase and 27.5 μM Biotinyl tyramide. 3. On ice, wash the cells three times with PBS. The bound Biotinyl tyramide is visualized by incubation with fluorescent streptavidin (1:1000, prepared in PBS) at 4°C for 30 min. 4. Fix the cells with ice-cold 4% paraformaldehyde solution and stain the nuclei. 5. Observe using a fluorescence microscope.

Biotinyl tyramide (100 μM, 5 min) and Hemin (HY-19424) enhanced biotinylation in starved U2OS cells after confirming the G4-specific biotinylation activity of the RNA-Hemin complex in vitro^[6].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

363.47

C₁₈H₂₅N₃O₃S

41994-02-9

Solid

White to off-white

O=C(NCCC1=CC=C(O)C=C1)CCCC[C@@H]2SC[C@]([C@]2([H])N3)([H])NC3=O

Room temperature in continental US; may vary elsewhere.

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Evans M F, et al. Optimization of biotinyl-tyramide-based in situ hybridization for sensitive background-free applications on formalin-fixed, paraffin-embedded tissue specimens [J]. BMC Clinical Pathology, 2003, 3: 1-17.  [Content Brief]

  [2]. Bobrow M N, et al. Catalyzed reporter deposition, a novel method of signal amplification application to immunoassays [J]. Journal of immunological methods, 1989, 125(1-2): 279-285.  [Content Brief]

  [3]. Kim S H, et al. An improved protocol of biotinylated tyramine-based immunohistochemistry minimizing nonspecific background staining [J]. Journal of Histochemistry & Cytochemistry, 2003, 51(1): 129-132.  [Content Brief]

  [4]. Hunyady B, et al. Immunohistochemical signal amplification by catalyzed reporter deposition and its application in double immunostaining [J]. Journal of Histochemistry & Cytochemistry, 1996, 44(12): 1353-1362.  [Content Brief]

  [5]. Dráberová E, et al. Quantification of α-tubulin isotypes by sandwich ELISA with signal amplification through biotinyl-tyramide or immuno-PCR [J]. Journal of immunological methods, 2013, 395(1-2): 63-70.  [Content Brief]

  [6]. Kharel P, et al. Stress promotes RNA G-quadruplex folding in human cells [J]. Nature Communications, 2023, 14(1): 205.  [Content Brief]

  [7]. Pató A, et al. Hydrogen peroxide production by epidermal dual oxidase 1 regulates nociceptive sensory signals [J]. Redox Biology, 2023, 62: 102670.  [Content Brief]