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Rnascope 2.5 hd detection reagent kit

Manufactured by Advanced Cell Diagnostics
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The RNAScope 2.5 HD Detection Reagent Kit is a laboratory equipment product designed for in situ hybridization (ISH) analysis. The kit provides reagents for the detection and visualization of target RNA molecules within tissue or cell samples.

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Lab products found in correlation

Hybez oven, by Advanced Cell Diagnostics (3 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Hematoxyline eosine, by Merck Group (1 mentions) Dm5000b microscope, by Leica (1 mentions) Hrp conjugated goat anti mouse igg, by Sangon (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Monospin c18, by GL Sciences (1 mentions) Hrp conjugated goat anti rabbit igg, by Sangon (1 mentions) Edta free protease inhibitor cocktail tablet, by Roche (1 mentions) Phosphatase inhibitor cocktail tablet, by Roche (1 mentions)

Close spelling variants of this product

RNAscope kit (59 citations) RNAscope 2.5 HD Detection Kit (27 citations) RNAscope 2.5 HD Reagent Kit (25 citations) RNAscope 2.5 HD detection reagents-RED kit (2 citations) RNAscope 2.5 HD Detection Reagents-BROWN kit (2 citations)

7 protocols using rnascope 2.5 hd detection reagent kit

1

In situ hybridization of Adamts6 and Adamts10

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Adamts6 and Adamts10 ISH was performed using RNAscope (Advanced Cell Diagnostics, Newark, CA) as described (Mead and Apte, 2020b (link)). Briefly, 6 μm sections were deparaffinized and hybridized to mouse Adamts6 and Adamts10 probe sets (428301, 585161, respectively; Advanced Cell Diagnostics) using a HybEZ oven (Advanced Cell Diagnostics) and the RNAScope 2.5 HD Detection Reagent Kit (322360; Advanced Cell Diagnostics) and counterstained with eosin.
Mead T.J., Martin D.R., Wang L.W., Cain S.A., Gulec C., Cahill E., Mauch J., Reinhardt D., Lo C., Baldock C, & Apte S.S. (2022). Proteolysis of fibrillin-2 microfibrils is essential for normal skeletal development. eLife, 11, e71142.
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2

Spatial RNA Expression in Mouse Knee

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Knees of 3‐month‐old mice were fixed in 4% paraformaldehyde for 48 hours at 4°C, demineralized in Morse’s solution (22.5% formic acid, 10 g/100 ml sodium citrate) for 48 hours at 4°C, embedded in paraffin, and sectioned sagittally at 7‐µm thickness. RNA in situ hybridization was performed by using specific probes (Supplementary Table 3) and RNAscope 2.5 HD Detection Reagent Kit (Advanced Cell Diagnostics). Pictures were taken by using an AxioScan.Z1 (Zeiss) instrument and processed by using Adobe Photoshop CC 19.1.6 software. Weak RNA signals were amplified by increasing the magenta color saturation, as indicated in Figures 3‐5.
Jones K., Angelozzi M., Gangishetti U., Haseeb A., de Charleroy C., Lefebvre V, & Bhattaram P. (2021). Human Adult Fibroblast‐like Synoviocytes and Articular Chondrocytes Exhibit Prominent Overlap in Their Transcriptomic Signatures. ACR Open Rheumatology, 3(6), 359-370.
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3

MAALIN Expression in Hippocampal Tissue

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Frozen fixed hippocampal tissue was probed with a MAALIN-specific probe and detected using RNAscope 2.5 HD Detection Reagent kit (Advanced Cell Diagnostics). Sections were counter-stained using hematoxyline-eosine (Sigma) and visualized using Leica DM 5000B microscope at 40X. Visual counts were used to determine the percentage of MAALIN-expressing cells.
Labonté B., Abdallah K., Maussion G., Yerko V., Yang J., Bittar T., Quessy F., Golden S.A., Navarro L., Checknita D., Gigek C., Lopez J.P., Neve R.L., Russo S.J., Tremblay R.E., Côté G., Meaney M.J., Mechawar N., Nestler E.J, & Turecki G. (2020). Regulation of impulsive and aggressive behaviours by a novel lncRNA. Molecular psychiatry, 26(8), 3751-3764.
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4

In Situ Hybridization of Adamts18

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Adamts18 ISH was performed using RNAScope (Advanced Cell Diagnostics, Newark, CA) following the manufacturer’s protocol. Briefly, 4 µm sections were deparaffinized and hybridized to a mouse Adamts18 probe set (452251; Advanced Cell Diagnostics) using a HybEZ oven (Advanced Cell Diagnostics) and the RNAScope 2.5 HD Detection Reagent Kit (322360; Advanced Cell Diagnostics) and stained with anti-SMA after the RNAScope procedure.
Ataca D., Aouad P., Constantin C., Laszlo C., Beleut M., Shamseddin M., Rajaram R.D., Jeitziner R., Mead T.J., Caikovski M., Bucher P., Ambrosini G., Apte S.S, & Brisken C. (2020). The secreted protease Adamts18 links hormone action to activation of the mammary stem cell niche. Nature Communications, 11, 1571.
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5

VCAN Expression Detection via Chromogenic FISH

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Chromogenic in situ hybridization for VCAN (Probe-Hs-VCAN, Cat No. 430071, Advanced Cell Diagnostics Inc. USA) was performed using the RNAscope 2.5 HD Detection Reagent kit (Advanced Cell Diagnostics Inc.) according to the manufacturer’s instructions (see Supplementary Methods).
Pearce O.M., Delaine-Smith R., Maniati E., Nichols S., Wang J., Böhm S., Rajeeve V., Ullah D., Chakravarty P., Jones R.R., Montfort A., Dowe T., Gribben J., Jones J.L., Kocher H.M., Serody J.S., Vincent B.G., Connelly J., Brenton J.D., Chelala C., Cutillas P.R., Lockley M., Bessant C., Knight M, & Balkwill F.R. (2017). Deconstruction of a metastatic tumor microenvironment reveals a common matrix response in human cancers. Cancer discovery, 8(3), 304-319.
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6

Paraformaldehyde Fixation and Immunostaining for Cx43 in Myocardium

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Tissues were fixed in 4% paraformaldehyde in PBS at 4 °C overnight followed by paraffin embedding. Sections of 7 μm were used for hematoxylin and eosin staining, picrosirius red staining, and immunofluorescence for Cx43 (catalog no. C6219; 1:800; Sigma-Aldrich) followed by secondary goat anti-rabbit antibody (catalog no. 111–035-144; 1:2000; Jackson Immunoresearch Laboratories Inc.). Antigen retrieval, i.e. immersion of slides in citrate-EDTA buffer (10 mM/L citric acid, 2 mM/L EDTA, 0.05% v/v Tween-20, pH 6.2) and microwaving for 1.5 min at 50% power four times in a microwave oven with 30-s intervals intervening was used before immunofluorescence. Immunofluorescence was quantified by the ratio of Cx43 signal to DAPI-positive cell nuclei integrated density (ImageJ; National Institutes of Health, n = 3, with three samples of each myocardium). Adamts6 RNA in situ hybridization was performed using RNAScope (Advanced Cell Diagnostics) following the manufacturer’s protocol. Briefly, 7-μm sections were deparaffinized and hybridized to a mouse Adamts6 probe set (catalog no. 428301; Advanced Cell Diagnostics) using a HybEZ™ oven (Advanced Cell Diagnostics) and the RNAScope 2.5 HD Detection Reagent Kit (catalog no. 322360; Advanced Cell Diagnostics).
Prins B.P., Mead T.J., Brody J.A., Sveinbjornsson G., Ntalla I., Bihlmeyer N.A., van den Berg M., Bork-Jensen J., Cappellani S., Van Duijvenboden S., Klena N.T., Gabriel G.C., Liu X., Gulec C., Grarup N., Haessler J., Hall L.M., Iorio A., Isaacs A., Li-Gao R., Lin H., Liu C.T., Lyytikäinen L.P., Marten J., Mei H., Müller-Nurasyid M., Orini M., Padmanabhan S., Radmanesh F., Ramirez J., Robino A., Schwartz M., van Setten J., Smith A.V., Verweij N., Warren H.R., Weiss S., Alonso A., Arnar D.O., Bots M.L., de Boer R.A., Dominiczak A.F., Eijgelsheim M., Ellinor P.T., Guo X., Felix S.B., Harris T.B., Hayward C., Heckbert S.R., Huang P.L., Jukema J.W., Kähönen M., Kors J.A., Lambiase P.D., Launer L.J., Li M., Linneberg A., Nelson C.P., Pedersen O., Perez M., Peters A., Polasek O., Psaty B.M., Raitakari O.T., Rice K.M., Rotter J.I., Sinner M.F., Soliman E.Z., Spector T.D., Strauch K., Thorsteinsdottir U., Tinker A., Trompet S., Uitterlinden A., Vaartjes I., van der Meer P., Völker U., Völzke H., Waldenberger M., Wilson J.G., Xie Z., Asselbergs F.W., Dörr M., van Duijn C.M., Gasparini P., Gudbjartsson D.F., Gudnason V., Hansen T., Kääb S., Kanters J.K., Kooperberg C., Lehtimäki T., Lin H.J., Lubitz S.A., Mook-Kanamori D.O., Conti F.J., Newton-Cheh C.H., Rosand J., Rudan I., Samani N.J., Sinagra G., Smith B.H., Holm H., Stricker B.H., Ulivi S., Sotoodehnia N., Apte S.S., van der Harst P., Stefansson K., Munroe P.B., Arking D.E., Lo C.W, & Jamshidi Y. (2018). Exome-chip meta-analysis identifies novel loci associated with cardiac conduction, including ADAMTS6. Genome Biology, 19, 87.
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7

Ketamine Pharmacological Assay Protocol

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Ketamine was dissolved in 0.9% saline for intraperitoneal (IP) injection (10 mg/kg). The 0.9% saline was used as the vehicle. EDTA-free protease inhibitor cocktail tablets (#05892791001) and Phosphatase inhibitor cocktail tablets (#04906837001) were purchased from Roche (Basel, Switzerland). Pierce™ BCA Protein Assay Kit (#23225) and anti-phospho-Csnk1a1 (#PA5-36790) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Titansphere Phos-TiO2 kit (#5010-21312) and Monospin C18 (#5010-21701) were purchased from GL Sciences (Japan). Anti-GAPDH (#sc-32233) was purchased from Santa Cruz (Dallas, TX, USA). HRP-conjugated Goat Anti-Rabbit IgG (#D110058) and HRP-conjugated Goat Anti-Mouse IgG (#D110087) were purchased from the Sangon Biotech (Shanghai, China). RNAscope 2.5 HD detection reagent kit (#322350) and Prkcg probe (#417911) were purchased from Advanced Cell Diagnostics (Newark, NJ, USA). Urea (U0631), ammonium bicarbonate (ABC) and other materials were obtained from Sigma–Aldrich (St. Louis, MO, USA) unless otherwise indicated.
Xiao Y., Luo H., Yang W.Z., Zeng Y., Shen Y., Ni X., Shi Z., Zhong J., Liang Z., Fu X., Tu H., Sun W., Shen W.L., Hu J, & Yang J. (2020). A Brain Signaling Framework for Stress-Induced Depression and Ketamine Treatment Elucidated by Phosphoproteomics. Frontiers in Cellular Neuroscience, 14, 48.
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