Alexa fluor tyramide signal amplification reagents

Manufactured by Thermo Fisher Scientific

Sourced in United States

Alexa Fluor Tyramide signal amplification reagents are a set of fluorescent detection reagents used in immunohistochemistry and in situ hybridization applications. They are designed to enhance the signal detection sensitivity of target biomolecules.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Benchmark ultra, by Roche (1 mentions) Zen software, by Zeiss (1 mentions) Sp8 inverted microscope, by Leica (1 mentions) Anti erα antibody d8h8, by Cell Signaling Technology (1 mentions) Ultraview dab detection kit, by Roche (1 mentions) Bond polymer refine detection kit, by Leica (1 mentions) Polymer refine detection kit, by Leica (1 mentions) Pannoramic confocal scanner, by 3DHISTECH (1 mentions)

Spelling variants of this product

Alexa Fluors (21 citations) Tyramide signal amplification (18 citations) Tyramide Reagent (2 citations)

2 protocols using alexa fluor tyramide signal amplification reagents

Estrogen Receptor Alpha Immunostaining

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ERα immunostaining (IHC) was performed on Benchmark Ultra using the ultraView DAB Detection kit (Ventana, USA). Antigen retrieval was performed onboard with UltraCC1 buffer (pH 8.2–8.5) at 95 °C for 52 min. Primary antibody (anti-ERα antibody D8H8, Cell Signaling Technology, USA, #8644) 1:100 for 28 min at 37 °C. Secondary antibody 1:100 for 1 h. Images were obtained using a Zeiss Axiovert Widefield Microscope and Zeiss ZEN software (Carl Zeiss, Germany). Immunofluorescence was performed using Leica Bond RX stainer and the Bond Polymer Refine Detection kit (Leica, Germany). Antibodies and detection: OXPHOS (2 µg/ml, Invitrogen, USA, #A-21347); ERα D8H8 (1 µg/ml, Cell Signaling Technology, USA, #8644); Alexa Fluor Tyramide signal amplification reagents (Life Technologies, USA); 4′, 6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich, USA). Slides were mounted in Mowiol 4–88 (Calbiochem, USA). Confocal imaging was performed on a Leica SP8 inverted microscope (Leica, Germany). Image processing and analysis (2-D and 3-D) was performed using Imaris software (Bitplane, CH).

Strillacci A., Sansone P., Rajasekhar V.K., Turkekul M., Boyko V., Meng F., Houck-Loomis B., Brown D., Berger M.F., Hendrickson R.C., Chang Q., de Stanchina E., Pareja F., Reis-Filho J.S., Rajappachetty R.S., Del Priore I., Liu B., Cai Y., Penson A., Mastroleo C., Berishaj M., Borsetti F., Spisni E., Lyden D., Chandarlapaty S, & Bromberg J. (2022). ERα-LBD, an isoform of estrogen receptor alpha, promotes breast cancer proliferation and endocrine resistance. NPJ Breast Cancer, 8, 96.

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Ferritin Immunofluorescence Detection Protocol

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The immunofluorescence (IF) detection of ferritin was performed at the Molecular Cytology Core Facility of Memorial Sloan Kettering Cancer Center. Automated IF staining was conducted using Leica Bond BX staining system. Cells cultured on chamber slides were fixed in 4 % paraformaldehyde (PFA, Electron Microscopy Sciences, 157–8–100) for 15 min followed by washing in PBS. Slides were loaded in Leica Bond and IF staining was performed as follows. The primary antibodies against FTH1 (Rb, 0.11μg/mL, Cell signaling technology, 4393) were incubated with the sample for 1 hour at room temperature. After that, the Leica Bond Polymer anti-rabbit HRP secondary antibody (included in Polymer Refine Detection Kit (Leica, DS9800)) was used for 8 min, followed by incubation with Alexa Fluor tyramide signal amplification reagents (Life Technologies, B40953) for 10 min. After the run was finished, slides were washed in PBS and incubated in 5 μg/mL 4’,6-diamidino-2-phenylindole (DAPI) (Sigma Aldrich, D9542) in PBS for 5 min, rinsed in PBS, and mounted in Mowiol 4–88 (Calbiochem). Slides were kept overnight at - 20°C before imaging. The slides were scanned with a 20x/0.8NA objective on a Pannoramic Confocal Scanner (3DHistech, Budapest, Hungary).

Richard N., Salomon H., Rando R., Mansour T., Bowlin T.L, & Wainberg M.A. (2000). Selection and Characterization of Human Immunodeficiency Virus Type 1 Variants Resistant to the (+) and (−) Enantiomers of 2′-Deoxy-3′-Oxa-4′-Thio-5-Fluorocytidine. Antimicrobial Agents and Chemotherapy, 44(5), 1127-1131.

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