Brightvision poly hrp linker

Manufactured by Immunologic

Brightvision poly-HRP-linker is a laboratory equipment designed to enable efficient and sensitive detection of target analytes in various immunoassay applications. It functions as a universal detection reagent that amplifies signal intensity, providing enhanced sensitivity for the identification and quantification of specific targets.

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Lab products found in correlation

Dab substrate, by Agilent Technologies (2 mentions) Anti mouse cy3, by Jackson ImmunoResearch (1 mentions) Ab72625, by Abcam (1 mentions) Z0458, by Agilent Technologies (1 mentions) Rabbit anti mouse hrp, by Agilent Technologies (1 mentions) Bx40 microscope, by Olympus (1 mentions)

Spelling variants of this product

Brightvision (5 citations) Brightvision-HRP (3 citations)

2 protocols using brightvision poly hrp linker

Immunohistochemical and Immunofluorescence Analysis

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Tissues were fixed overnight in 4% paraformaldehyde and embedded in paraffin according to standard protocols. Sections (6μm) were deparaffinized followed by antigen retrieval using microwave treatment in 0.01M sodium citrate. Endogenous peroxidase activity was blocked and immunostaining was performed overnight at 4°C using TR2A (Abcam, ab72625) and 8FM 1:10 antibodies (Buijsen et al., 2014 (link)). In order to better visualize inclusions an extra antigen retrieval step was added, using proteinase K. Antigen-antibody complexes were visualized by incubation with DAB substrate (Dako, K3468) after incubation with Brightvision poly-HRP-linker (Immunologic, DPVO-HRP 55). Slides were counterstained with hematoxylin and mounted with Entellan.
For (double) immunofluorescence, slides were blocked for auto-fluorescence with Sudan Black in 70% ethanol. Primary antibodies include TR2A (Abcam, ab72625), 8FM 1:10 (Buijsen et al., 2014 (link)) and ubiquitin (Dako, Z0458). Secondary antibodies were antirabbit Fab 488 (Molecular Probes, A11070) and antimouse cy3 (Jackson, 715-165-150). Nuclei were visualized with Hoechst (Figure S7).

Cid-Samper F., Gelabert-Baldrich M., Lang B., Lorenzo-Gotor N., Ponti R.D., Severijnen L.A., Bolognesi B., Gelpi E., Hukema R.K., Botta-Orfila T, & Tartaglia G.G. (2018). An Integrative Study of Protein-RNA Condensates Identifies Scaffolding RNAs and Reveals Players in Fragile X-Associated Tremor/Ataxia Syndrome. Cell Reports, 25(12), 3422-3434.e7.

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Immunohistochemical Analysis of Human Brain

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Human brain sections (6 μm) were deparaffinized in xylene and rehydrated (100%–96%-90%–80%-70%–50% EtOH serie). Antigen retrieval was done in 0.01 M sodium citrate, pH 6.0 using pressure cooker treatment. Endogenous peroxidase activity was blocked with 0.6% H₂O₂ and 1,25% sodium azide in 0.1 M PBS. Immunostaining was performed overnight at 4 °C in PSB block buffer (0.1 M PBS/0.5%protifar/0.15%glycine). Antibodies used in this study are listed in Additional file 1: Table S3, including concentration and brand/catalogue number. Antigen-antibody complexes were visualized by incubation with DAB substrate (DAKO) after incubation with Brightvision poly-HRP-linker (Immunologic) or anti-mouse/rabbit HRP (DAKO). Slides were counterstained with Mayer’s haematoxylin and mounted with Entellan (Merck Millipore International). The slides were then left to dry in the fume hood for an hour and thereafter put in an 37 °C incubater overnight. Pictures were taken by using an Olympus BX40 microscope (Olympus).

Riemslagh F.W., Lans H., Seelaar H., Severijnen L.A., Melhem S., Vermeulen W., Aronica E., Pasterkamp R.J., van Swieten J.C, & Willemsen R. (2019). HR23B pathology preferentially co-localizes with p62, pTDP-43 and poly-GA in C9ORF72-linked frontotemporal dementia and amyotrophic lateral sclerosis. Acta Neuropathologica Communications, 7, 39.

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