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Dako aqueous mounting medium

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Dako aqueous mounting medium is a laboratory reagent used to mount and preserve samples for microscopic analysis. It is a water-based solution that helps maintain the structural integrity and clarity of specimens during the mounting process.

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Lab products found in correlation

Sp8 upright confocal microscope, by Leica camera (2 mentions) Alexa fluor 633 goat anti mouse igg h l, by Thermo Fisher Scientific (2 mentions) Anti digoxigenin antibody, by Roche (1 mentions) Nbt bcip, by Promega (1 mentions) Digoxigenin rna labeling mix, by Roche (1 mentions) T7 rna polymerase, by Promega (1 mentions) Superfrost plus, by Thermo Fisher Scientific (1 mentions) Pgem t vector system, by Promega (1 mentions)

3 protocols using dako aqueous mounting medium

1

Immunofluorescent Staining of Mouse Brain

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After euthanasia, the mouse was perfused with 0.25 mg/mL heparin in ice-cold PBS. Then, the mouse was perfused with ice-cold 4% paraformaldehyde (PFA). The brain was dissected and transferred to 4% PFA for 2 hours on ice. The brains were transferred to 20% sucrose in PBS and incubated overnight at 4°C. The next day, the brains were transferred to 30% sucrose in PBS and incubated overnight at 4°C. The brains were embedded in OCT and 10 μm sections were cut. The brain sections were washed 2× in TBS and blocked in 5% normal goat serum in TBST for 1 hour at room temperature. The brains were stained overnight at 4°C with HER2 (Santa Cruz sc-33684, 1:250). The sections were washed 3× with TBST and incubated with secondary antibody Alexa Fluor 633 Goat Anti Mouse IgG (H + L) (Thermo Fisher, 1:250) for 1 hour in the dark at room temperature. The slides were washed 3× with TBST. The sections were stained with Hoescht 33342 (5 μg/mL) for 10 minutes at room temperature in the dark. The sections were washed 3× with TBST and 1× with TBS. The cover slips were mounted using Dako aqueous mounting medium (Dako). The sections were imaged using a Leica SP8 upright confocal microscope.
McKernan C.M., Khatri A., Hannigan M., Child J., Chen Q., Mayro B., Snyder D., Nicchitta C.V, & Pendergast A.M. (2022). ABL kinases regulate translation in HER2+ cells through Y-box-binding protein 1 to facilitate colonization of the brain. Cell reports, 40(9), 111268.
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2

Immunofluorescent Staining of Mouse Brain

Check if the same lab product or an alternative is used in the 5 most similar protocols
After euthanasia, the mouse was perfused with 0.25 mg/mL heparin in ice-cold PBS. Then, the mouse was perfused with ice-cold 4% paraformaldehyde (PFA). The brain was dissected and transferred to 4% PFA for 2 hours on ice. The brains were transferred to 20% sucrose in PBS and incubated overnight at 4°C. The next day, the brains were transferred to 30% sucrose in PBS and incubated overnight at 4°C. The brains were embedded in OCT and 10 μm sections were cut. The brain sections were washed 2× in TBS and blocked in 5% normal goat serum in TBST for 1 hour at room temperature. The brains were stained overnight at 4°C with HER2 (Santa Cruz sc-33684, 1:250). The sections were washed 3× with TBST and incubated with secondary antibody Alexa Fluor 633 Goat Anti Mouse IgG (H + L) (Thermo Fisher, 1:250) for 1 hour in the dark at room temperature. The slides were washed 3× with TBST. The sections were stained with Hoescht 33342 (5 μg/mL) for 10 minutes at room temperature in the dark. The sections were washed 3× with TBST and 1× with TBS. The cover slips were mounted using Dako aqueous mounting medium (Dako). The sections were imaged using a Leica SP8 upright confocal microscope.
McKernan C.M., Khatri A., Hannigan M., Child J., Chen Q., Mayro B., Snyder D., Nicchitta C.V, & Pendergast A.M. (2022). ABL kinases regulate translation in HER2+ cells through Y-box-binding protein 1 to facilitate colonization of the brain. Cell reports, 40(9), 111268.
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3

In Situ Hybridization for LRRK2 mRNA

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A probe for in situ hybridization was obtained by subcloning of a 500bp PCR fragment from hLRRK2 cDNA in pGEM-T vector system (Promega, Madison, Wi). Brains were rapidly removed, and frozen in OCT (Tissue-Tek) on dry ice. Sections (14 μm) were thaw-mounted onto glass slides (Superfrost Plus; Fisher Scientific). For hybridization, sections were warmed to 37°C and then fixed by immersion in 4% paraformaldehyde (PF) in 0.1 M phosphate buffer (PB) pH 7.1. Sections were then treated with a prehybridization solution as previously described (Burke et al., 1994 (link)). Sections were then covered with hybridization solution and incubated overnight at 68°C. Hybridization solution contained antisense hLRRK2 riboprobe obtained with digoxigenin RNA Labeling Mix (Roche, Mannheim, Germany) and T7 RNA polymerase (Promega). After washes, sections were incubated with an anti-digoxigenin antibody (Roche Diagnostics) at 1:5000 overnight at 4°C. After additional washes, sections were then incubated with BCIP/NBT (Promega, Madison WI) developing solution overnight at room temperature. Sections were washed and coverslipped with Dako aqueous mounting medium (Dako, CA).
Tagliaferro P., Kareva T., Oo T.F., Yarygina O., Kholodilov N, & Burke R.E. (2015). An Early Axonopathy in a hLRRK2(R1441G) Transgenic Model of Parkinson Disease. Neurobiology of disease, 82, 359-371.
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