Tsa plus cy3

Manufactured by PerkinElmer

Sourced in United States

The TSA Plus Cy3 is a laboratory equipment product designed for biomarker detection and amplification. It utilizes a proprietary signal amplification technology to enhance the detection of target analytes. The core function of the TSA Plus Cy3 is to provide a sensitive and reliable method for the visualization and analysis of biomolecules in various research and diagnostic applications.

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Lab products found in correlation

Lsm 700, by Zeiss (4 mentions) Rnascope assay, by Advanced Cell Diagnostics (3 mentions) Rnascope multiplex fluorescent v2 assay, by Bio-Techne (2 mentions) Mm ppib c1, by Bio-Techne (2 mentions) Dapb c1, by Bio-Techne (2 mentions) Fluoromountg, by BioConcept (2 mentions) Mm gpbar1 c1, by Bio-Techne (2 mentions) Mm agrp c2, by Bio-Techne (2 mentions) Withfluoromountg, by BioConcept (2 mentions) Goat anti gfp primary antibody, by Abcam (2 mentions) Donkey anti goatalexa488 secondary antibody, by Thermo Fisher Scientific (2 mentions) Lsm 700 confocal microscope, by Zeiss (2 mentions) Digoxigenin 11 utp, by Roche (1 mentions) Anti dig pod, by Roche (1 mentions) Anti dnp, by PerkinElmer (1 mentions) Clone sp2, by Thermo Fisher Scientific (1 mentions) Alexa 568 conjugated goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Ds fi2 camera, by Nikon (1 mentions) C2 confocal microscope, by Nikon (1 mentions)

8 protocols using tsa plus cy3

Multiplex RNAscope Fluorescent Imaging of Metabolic Genes

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RNAscope Multiplex Fluorescent V2 assay (Bio-techne, Cat#:323110) was
performed according to manufacturer’s instruction on 4 μm paraffin
sections, hybridized with the probes Mm-Gpbar1-C1(Bio-techne, Cat#:318451),
Mm-Agrp-C2 (Bio-techne, Cat#:400711-C2), Mm-Ppib-C1 (Bio-techne, Cat#:313911) as
positive control and DapB-C1 (Bio-techne, Cat#:310043) as negative control at
40°C for 2 hours and revealed with TSA Plus Cy3 (Perkin Elmer,
Cat#:NEL744E001KT). Tissues were counterstained with DAPI and mounted with
FluoromountG (Bioconcept, Cat#:100.01). When indicated, sections were incubated
overnight at 4°C with a goat anti-GFP primary antibody (Abcam,
Cat#:ab6673 – 1:200 dilution). After incubation with a donkey anti-goat
Alexa488 secondary antibody (Thermo Fisher, Cat#:A11055, 1:1000 dilution),
tissues were counterstained with DAPI and mounted with FluoromountG (Bioconcept,
Cat#:100.01). The imaging was performed with Zeiss LSM 700 confocal microscope
(Carl Zeiss Microscopy, Germany).

Perino A., Velázquez-Villegas L.A., Bresciani N., Sun Y., Huang Q., Fénelon V.S., Castellanos-Jankiewicz A., Zizzari P., Bruschetta G., Jin S., Baleisyte A., Gioiello A., Pellicciari R., Ivanisevic J., Schneider B.L., Diano S., Cota D, & Schoonjans K. (2021). Central anorexigenic actions of bile acids are mediated by TGR5. Nature metabolism, 3(5), 595-603.

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In Situ Hybridization Protocols for Diverse Organisms

Cited 2 times

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Animals were manually collected, fixed, and processed for in situ hybridization as described [79 (link),80 (link)]. Labeled antisense RNA probes were transcribed from linearized DNA using digoxigenin-11-UTP (Roche, Basel, Switzerland) or labeled with DNP (Mirus Bio, Madison, WI, USA) according to the manufacturer’s instructions. For I. pulchra and M. stichopi, colorimetric WMISH was performed according to the protocol outlined in [79 (link)]. For N. vectensis, we followed the protocol described by [81 ]. Double fluorescent in situ hybridization (FISH) was performed as the colorimetric WMISH with the following modifications: after the posthybridization steps, animals were incubated overnight with peroxidase-conjugated antibodies at 4 °C (anti-DIG-POD [Roche, Basel, Switzerland], 1:500 dilution, and anti-DNP [Perkin Elmer, Waltham, MA, USA], 1:200 dilution) followed by the amplification of the signal with fluorophore-conjugated tyramides (1:100 TSA reagent diluents [Perkin Elmer, Waltham, MA, USA] TSA Plus Cy3 or Cy5 Kit). Residual enzyme activity was inhibited via 45-minute incubation in 0.1% hydrogen peroxide in PTW followed by four PTW washes prior to addition and development of the second peroxidase-conjugated antibody [82 (link)].

Andrikou C., Thiel D., Ruiz-Santiesteban J.A, & Hejnol A. (2019). Active mode of excretion across digestive tissues predates the origin of excretory organs. PLoS Biology, 17(7), e3000408.

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Spatial Transcriptomics of RANKL and WNT4

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RNAscope assay (Advanced Cell Diagnostics, Cat. No. 323110) was performed according to manufacturer’s protocol on 4 μm deparaffinized sections and with probes for RANKL (ACD, Cat No. 523331‐C2), WNT4 (ACD, Cat No. 429441), Mm‐Ppib (ACD, Cat. No. 313911, positive control), and DapB (ACD, Cat. No. 310043, negative control) at 40°C for 2 h and revealed with TSA Plus‐Cy3 (Perkin Elmer, Cat. No. NEL744001KT). WNT4 and RANKL were revealed with Opal570 and Opal650, respectively. Confocal images were acquired using a laser‐scanning confocal inverted microscope (LSM700, Carl Zeiss, Inc.).

Shamseddin M., De Martino F., Constantin C., Scabia V., Lancelot A., Laszlo C., Ayyannan A., Battista L., Raffoul W., Gailloud‐Matthieu M., Bucher P., Fiche M., Ambrosini G., Sflomos G, & Brisken C. (2021). Contraceptive progestins with androgenic properties stimulate breast epithelial cell proliferation. EMBO Molecular Medicine, 13(7), e14314.

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Detecting LOXL1 Expression with RNAscope

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RNAscope assay (Advanced Cell Diagnostics, Cat. No. 323110) was performed according to manufacturer's protocol on 4 μm deparaffinized sections using the following probes: Hs‐LOXL1 (Cat No. 470751), Mm‐Ppib (ACD, Cat. No. 313911, positive control) and DapB (ACD, Cat. No. 310043, negative control) at 40°C for 2 h and revealed with TSA Plus‐Cy3 (Perkin Elmer, Cat. No. NEL744001KT). Images were captured on confocal Zeiss LSM700.

Sflomos G., Battista L., Aouad P., De Martino F., Scabia V., Stravodimou A., Ayyanan A., Ifticene‐Treboux A., Bucher P., Fiche M., Ambrosini G, & Brisken C. (2021). Intraductal xenografts show lobular carcinoma cells rely on their own extracellular matrix and LOXL1. EMBO Molecular Medicine, 13(3), e13180.

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Multiplex RNAscope and Immunofluorescence

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RNAscope assay (Advanced Cell Diagnostics, Cat. No. 323110) was performed according to manufacturer’s protocol on 4 μm deparaffinized sections and hybridised with probes: Mm-Esr1 (ACD, Cat. No. 432861), Mm-Areg (ACD, Cat. No. 430501), Mm-Ppib (ACD, Cat. No. 313911, positive control) and DapB (ACD, Cat. No. 310043, negative control) at 40 °C for 2h and revealed with TSA Plus-Cy3 (Perkin Elmer, Cat. No. NEL744001KT). Rabbit anti-PgR (1:400, clone SP2, Thermo Fisher, Cat. No.: RM-9102-P) and rabbit anti-ERα (1:100; MC20, sc-542 SantaCruz) was incubated overnight at 4 °C and detected with Alexa 488 or Alexa 568 conjugated goat anti-rabbit (1:1000, Life Technology), respectively. Images were captured on confocal Zeiss LSM700 and spots quantified using QuPath and an in-house script, code available from O. Burri at the Bioimaging and Optics platform, EPFL, based on the guide for RNAscope Data Analysis.

Cagnet S., Ataca D., Sflomos G., Aouad P., Schuepbach-Mallepell S., Hugues H., Krust A., Ayyanan A., Scabia V, & Brisken C. (2018). Oestrogen receptor α AF-1 and AF-2 domains have cell population-specific functions in the mammary epithelium. Nature Communications, 9, 4723.

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Zebrafish Whole Mount In Situ Hybridization

Cited 5 times

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Whole mount in situ hybridization (WISH) was conducted as described^{101 (link),102 (link)}. Anti-sense RNA probes were digoxigenin-labeled (etv5a, odf3b, cetn4, scl, irx2a, cdh17, slc20a1a, trmp7, slc12a1, slc12a3, jag2b) or fluorescein-labeled (smyhc1, cdh17, pax2a), and generated by in vitro transcription using plasmid or PCR templates as described^{16 (link),17 (link),36 (link),63 (link),103 (link)}. Embryos were mounted in glycerol and images were taken using a Nikon Eclipse Ni with a DS-Fi2 camera. Whole mount fluorescent in situ hybridization (FISH) was performed as described^{103 (link)} with the digoxigenin and fluorescein-labeled RNA probes used in WISH. Stains were developed with the TSA plus Cy3 and TSA plus Fluorescein kits (Perkin Elmer). Embryos were mounted in Poly Aqua-mount as described^{104 (link)} and imaged on a Nikon C2 confocal microscope. Z-stacks were processed with FIJI into max image projections, and all figures were assembled using Adobe Photoshop CS5.

Marra A.N., Cheng C.N., Adeeb B., Addiego A., Wesselman H.M., Chambers B.E., Chambers J.M, & Wingert R.A. (2019). Iroquois transcription factor irx2a is required for multiciliated and transporter cell fate decisions during zebrafish pronephros development. Scientific Reports, 9, 6454.

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Multiplex RNAscope Fluorescent Imaging of Metabolic Genes

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Fluorescent In Situ Hybridization Protocol

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For fluorescent ISH, cells were rinsed in PBS twice, fixed in 4% formaldehyde in PBS (pH 7.4) for 30 min at room temperature, incubated in a H₂O₂ solution for 10 min and digested in protease solution for 10 min. The slides were then hybridized with a custom probe, Hs-LOC105372310-O2, in a HybEZ oven (Advanced Cell Diagnostics) at 40 °C for 2 h. After signal amplification, the slides were conjugated with TSA® Plus Cy3 (PerkinElmer). The slides were then incubated with anti-NKRF antibody overnight at 4 °C. After incubation with the fluorescein-conjugated secondary antibody, the slides were mounted with ProLong® Gold Antifade Reagent with DAPI.

Jin X., Ge L.P., Li D.Q., Shao Z.M., Di G.H., Xu X.E, & Jiang Y.Z. (2020). LncRNA TROJAN promotes proliferation and resistance to CDK4/6 inhibitor via CDK2 transcriptional activation in ER+ breast cancer. Molecular Cancer, 19, 87.

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