Biotinyl tyramide

Manufactured by PerkinElmer

Sourced in United States

Biotinyl tyramide is a reagent used in various biotechnology applications, including signal amplification in immunoassays and in situ hybridization techniques. It is a small molecule that can be enzymatically incorporated into target biomolecules, allowing for the detection and visualization of these targets.

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Lab products found in correlation

Extra avidin fitc, by Merck Group (2 mentions) Extra avidin tritc, by Merck Group (2 mentions) Lsm 710, by Zeiss (2 mentions) Abc reagent, by Thermo Fisher Scientific (1 mentions) Fluoromount mounting medium with dapi, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 fluorescent secondary antibody, by Thermo Fisher Scientific (1 mentions) Rabbit anti gfp antibody, by Abcam (1 mentions) Vectastain elite abc reagent, by Vector Laboratories (1 mentions) Ab5026, by Merck Group (1 mentions) Histofine simple stain kit, by Nichirei Biosciences (1 mentions) Biotinyl tyramide stock solution, by PerkinElmer (1 mentions) Plus amplification diluent, by PerkinElmer (1 mentions) Malinol, by Muto Pure Chemicals (1 mentions) Tsa biotin system, by PerkinElmer (1 mentions) Fluoromount g mounting medium, by Southern Biotech (1 mentions) Streptavidin hrp, by Jackson ImmunoResearch (1 mentions) Rabbit anti cgrp, by Merck Group (1 mentions) Abc reagent, by Vector Laboratories (1 mentions) Rabbit anti pkcγ, by Santa Cruz Biotechnology (1 mentions) Rabbit anti serotonin, by Merck Group (1 mentions)

Close spelling variants of this product

Biotinyl tyramide solution (2 citations) Biotinyl Tyramide Stock Solution (2 citations)

7 protocols using biotinyl tyramide

Chondroitin Sulfate Degradation Visualization

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To confirm successful intraspinal delivery of LV‐ChABC, sections spanning the injury epicenter were stained for C‐4‐S using tyramide signal amplification, to reveal stub epitopes which are only present after degradation of chondroitin sulfate glycosaminoglycans (CS‐GAGs). Sections were rehydrated with PBS and incubated at room temperature in the following: hydrogen peroxide (0.3%, 20 min), mouse monoclonal anti‐C‐4‐S (MP Biomedicals; 1:5000, overnight), biotinylated secondary antibody (anti‐mouse biotin; Vector Laboratories, USA; 1:400, 2 h), ABC reagent (ThermoFisher Scientific, USA; 1:250, 30 min), biotinyl tyramide (PerkinElmer Life Sciences, Boston, MA; 1:75, 10 min), and extra‐avidin FITC (Sigma‐Aldrich; 1:500, 3 h). Slides were coverslipped using Fluoromount mounting medium with DAPI (#CO‐4959‐52, Invitrogen). Representative images of C‐4‐S immunostaining for each treatment group were obtained using a confocal microscope (Zeiss, LSM710), using identical exposure and settings.

Sinopoulou E., Spejo A.B., Roopnarine N., Burnside E.R., Bartus K., De Winter F., McMahon S.B, & Bradbury E.J. (2022). Chronic muscle recordings reveal recovery of forelimb function in spinal injured female rats after cortical epidural stimulation combined with rehabilitation and chondroitinase ABC. Journal of Neuroscience Research, 100(11), 2055-2076.

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Visualizing Axonal Projections and GFP Signals

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To amplify and visualize GFP signals, 35-μm thick brain sections were incubated with a rabbit anti-GFP antibody (1:2000, Abcam). Signal was detected with a mouse Alexa Fluor 488 fluorescent secondary antibody (1:1000, Thermo Fisher Scientific). To visualize the CST projection fibers, 35-μm transverse spinal cord sections (C5–C8) were prepared from mice that received the BDA tracing. Solution containing streptavidin-horseradish peroxidase (Vectastain R.T.U. Elite ABC Reagent) was added to the sections for 1 hour followed by a 30 min incubation with PerkinElmer Biotinyl tyramide (1:100 in amplification diluent). Signals were detected following a 1-hour incubation with Extra-Avidin@ TRITC (1:200, Sigma-Aldrich).

Huang N., Li S., Xie Y., Han Q., Xu X.M, & Sheng Z.H. (2021). Reprogramming an energetic AKT-PAK5 axis boosts axon energy supply and facilitates neuron survival and regeneration after injury and ischemia. Current biology : CB, 31(14), 3098-3114.e7.

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Immunostaining Protocol for Methionine-Enkephalin

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The sections were incubated with rabbit polyclonal antibody against MEnk (AB5026 from Millipore; 1:50,000, 1:500,000, or 1:5,000,000) or without the anti-MEnk antibody for 18 h in PBS-BSA. After several rinses in PBS, the sections were incubated with the polymer staining reagent (Histofine Simple Stain Kit; Nichirei, Tokyo, Japan) for 30 min. After several rinses in PBS, they were processed for tyramide signal amplification (TSA) using the TSA Biotin System (Perkin Elmer, Boston, MA, USA). The sections were incubated in Biotinyl Tyramide (amplification reagent) Working Solution that was made by diluting Biotinyl Tyramide Stock Solution (Perkin Elmer) 1:50 using 1X Plus Amplification Diluent (Perkin Elmer, FP1135) for 30 min. After several rinses in PBS, the sections were incubated with the Vectastain Elite ABC reagent (Vector) for 30 min in PBS. After several rinses in PBS, the bound peroxidase was visualized by incubating the sections with a solution containing 0.05% DAB and 0.01% H₂O₂ in 0.05M Tris-HCl (pH 7.4) for 10 min. After several rinses in water, the immunostained sections were dehydrated and cover-slipped with Malinol (Muto Pure Chemicals). Overview protocol for the PBTA-DAB staining is shown in Figure 1 (left).

Goto S., Morigaki R., Okita S., Nagahiro S, & Kaji R. (2015). Development of a highly sensitive immunohistochemical method to detect neurochemical molecules in formalin-fixed and paraffin-embedded tissues from autopsied human brains. Frontiers in Neuroanatomy, 9, 22.

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Immunohistochemical Tyramide Amplification

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The 30-µm-thick sections were incubated with avidin-biotin-peroxidase (Vectastain R.T.U. Elite ABC Reagent, SAT700, PerkinElmer, USA) for 1 h. Biotinyl tyramide (PerkinElmer) was diluted to 1:100 with amplification diluent, then placed on the sections for an additional hour after the wash. The sections were then incubated with Extra-Avidin@ TRITC (Sigma, USA, 1:200 in 0.3% PBST) for 2 h. Lastly, the sections were washed again and coverslipped with SouthernBiotech Fluoromount-G mounting medium (Cat. No. 0100-01, SouthernBiotech).

Han Q., Ordaz J.D., Liu N.K., Richardson Z., Wu W., Xia Y., Qu W., Wang Y., Dai H., Zhang Y.P., Shields C.B., Smith G.M, & Xu X.M. (2019). Descending motor circuitry required for NT-3 mediated locomotor recovery after spinal cord injury in mice. Nature Communications, 10, 5815.

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Quantifying soluble PD-L1 variants

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To assay sPD-L1 variants, 0.1 µg/well of mouse anti-human PD-L1 mab (130021, R&D systems) or 0.2 µg/well anti-human PD-L1 mAb (29E.12B1) were coated on Costar ELISA plates overnight at 4°C. Plates were then washed with PBS and blocked with protein-free blocking buffer (Pierce, Rockford, IL) for 4 hours. Patient sera or plasma were diluted with PBS in 1:1 volume ratio. 100 µl per well of diluted patient sera or plasma were added and incubated overnight at 4°C. Plates were washed with PBS containing Tween-20, and incubated with 100 µl per well of 0.1µg/well biotinylated anti-PD-L1 mAb (29E.2A3, Biolegend) in protein-free blocking buffer at room temperature for 2 hours. Plates were then washed and incubated with 1mg/ml streptavidin-HRP (Jackson ImmunoResearch) diluted 1:40,000 in protein-free blocking buffer for 2 hours. Plates were washed and treated with biotinyl tyramide (Perkin Elmer) for 30 min, and then washed and incubated with 1 mg/ml streptavidin-HRP (Jackson ImmunoResearch) diluted 1:400,000 in protein-free blocking buffer for 2 hours with further development with TMB (Pierce). Plates were read at an optical density (O.D.) of 450 nm. All samples were assayed in duplicate. A standard curve using recombinant human PD-L1-HIS (Novoprotein, Summit, NJ) was also performed with each assay.

Zhou J., Mahoney K.M., Giobbie-Hurder A., Zhao F., Lee S., Liao X., Rodig S., Li J., Wu X., Butterfield L.H., Piesche M., Manos M.P., Eastman L.M., Dranoff G., Freeman G.J, & Hodi F.S. (2017). Soluble PD-L1 as a biomarker in malignant melanoma treated with checkpoint blockade. Cancer immunology research, 5(6), 480-492.

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Immunohistochemical Staining of Spinal Cord

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BDA staining was as follows: sections were incubated in 0.3% H
₂O
₂and 10% methanol (30 min). Sections were incubated in ABC reagent (VectorLabs) (30 min) then amplified using biotinyl tyramide (1:75, PerkinElmer), then left overnight at 4°C with extra avidin FITC (1:500, Sigma). Sections were washed between steps using PBS.
Series of 40-μm thick transverse sections of fixed spinal cord were immunolabelled as previously described (
Soleman
et al., 2010
). Primary antibodies (overnight) were: rabbit anti-PKCγ (Santa Cruz Biotechnology, 1:500); rabbit anti-CGRP (1:4000, Sigma); rabbit anti-serotonin (1:6000, Sigma). Secondary antibodies (3 h) were: donkey anti-rabbit IgG Alexa 546 (1:1000, Jackson Labs); goat anti-rabbit IgG Alexa 488 (1:1000, Sigma); goat anti-rabbit IgG Alexa 546 (1:1000, Sigma), and DAPI (1:50 000, Sigma). Sections were washed with PBS and then mounted and cover slipped with Mowiol
^®.

Duricki D.A., Hutson T.H., Kathe C., Soleman S., Gonzalez-Carter D., Petruska J.C., Shine H.D., Chen Q., Wood T.C., Bernanos M., Cash D., Williams S.C., Gage F.H, & Moon L.D. (2015). Delayed intramuscular human neurotrophin-3 improves recovery in adult and elderly rats after stroke. Brain, 139(1), 259-275.

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Articular Cartilage Morphological Analysis

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At the end of each culturing period (2, 4 or 6 weeks), a portion of the specimens were processed for morphology, morphometry of cell sizes, quantification of metachromasia (sulfated proteoglycans; internal normalization being performed by referring to the staining intensity of non-OA articular cartilage areas of the same joint) after staining with Toluidine Blue O, and for the immunhistochemical demonstration of type-II collagen.
The volume fraction of metachromasia was determined from the light micrographs by the point-counting technique, which was applied in accordance with stereological principles 26, 27 . The mean cell volumes were estimated using the point sampled intercept method 28 and systematic random sampling strategies 26 .
For the immunhistochemical demonstration of type-II collagen, the sections were first exposed to hyaluronidase, then to a type-II collagen antibody (clone CII C1; Hybridoma Bank, Iowa City, IA, USA). Immunoreactivity was enhanced by applying first the avidinbiotin-peroxidase complex (Vector Laboratories) and then biotinyl tyramide (Perkin Elmer, Waltham, MA, USA). Cell nuclei were counterstained with haematoxylin. The sections were evaluated and photographed in a Nikon Eclipse E1000 light microscope; for details see 14, 16

Hunziker E.B., Shintani N., Haspl M., Lippuner K., Vögelin E, & Keel M.J.B. (2022). The Synovium of Human Osteoarthritic Joints Retains Its Chondrogenic Potential Irrespective of Age. Tissue engineering. Part A, 28(5-6).

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