Basescope detection reagent kit v2 red

Manufactured by Advanced Cell Diagnostics

Sourced in United States

The BaseScope Detection Reagent Kit v2–RED is a set of reagents designed to enable the detection of target RNA molecules in tissue samples. The kit provides the necessary components for the hybridization, signal amplification, and chromogenic detection of specific RNA transcripts, allowing for the visualization and localization of gene expression patterns in various cell types and tissues.

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Lab products found in correlation

Superfrost plus slide, by Thermo Fisher Scientific (2 mentions) Hybez oven, by Advanced Cell Diagnostics (1 mentions) Rnascope protease 4, by Advanced Cell Diagnostics (1 mentions) Vectamount, by Vector Laboratories (1 mentions) Basescope, by Advanced Cell Diagnostics (1 mentions) Ecomount, by Biocare Medical (1 mentions) Rnascope multiplex fluorescent reagent kit v2, by Advanced Cell Diagnostics (1 mentions) Rnascope probe, by Advanced Cell Diagnostics (1 mentions) Prolong gold antifade mountant, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

BaseScope™ Reagent Kit v2-RED (9 citations) BaseScope (8 citations) BaseScope Red reagent kit (7 citations) BaseScope Reagent Kit (4 citations) BaseScope™ v2 Detection Reagent Kit (2 citations)

6 protocols using basescope detection reagent kit v2 red

In Situ Hybridization for Stra8 and Rara mRNA

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For detection of Stra8 mRNA, ISH was performed with digoxigenin-labeled probes as previously described (12 (link)). For detection of Rara mRNA, a commercially available kit was used, according to the manufacturer’s instructions (BaseScope Detection Reagent Kit v2–RED, Advanced Cell Diagnostics, 323900). Briefly, deparaffinized sections from PFA-fixed E11.5 embryos were treated with hydrogen peroxide for 10 min, washed, and boiled at 100°C in 1X Target Retrieval Reagent for 10 min. Next, Protease IV was then applied for 15 min at 40°C on dehydrated sections, and then, slides were washed in distilled water. The prewarmed probes (Rara and Ppib housekeeping gene positive control) were applied on the sections for 2 hours at 40°C. The slides were washed in 1X wash buffer and were subjected to a series of signal amplification (AMP1 to AMP8). Hybridization signals were detected using a chromogenic Fast RED-B/Fast RED-A reagent. The presence of Rara and Ppib mRNA was identified as red punctate dots. The sections were counterstained for 3 min with 12.5% (v/v) Harris hematoxylin diluted in distilled water.

Vernet N., Condrea D., Mayere C., Féret B., Klopfenstein M., Magnant W., Alunni V., Teletin M., Souali-Crespo S., Nef S., Mark M, & Ghyselinck N.B. (2020). Meiosis occurs normally in the fetal ovary of mice lacking all retinoic acid receptors. Science Advances, 6(21), eaaz1139.

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Gsdmc2-3 and Gsdmc4 Expression Analysis by BaseScope

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Intestinal tissue was processed for in situ hybridization using BaseScope™ Detection Reagent Kit v2-RED (323900, Advanced Cell Diagnostics) according to the manufacturer’s protocol. The test BaseScope probes 1) BA-Mm-Gsdmc2–3-zz and 2) BA-Mm-Gsdmc4-zz were ‘ZZ’ antisense probes designed to targeting mouse Gsdmc2–3 and Gsdmc4, respectively. The positive control probe is BA-Mm-Ppib-3zz and negative control probe: BA-Dapb-3zz.
Briefly, deparaffinized and dried sections were incubated with RNAscope® Hydrogen Peroxide for 10 min at room temperature and then 1xRNAscope® Target Retrieval Reagents for 15 min at 98~102°C. The samples were then incubated with RNAscope® Protease IV in HybEZ™ Oven (Advanced Cell Diagnostics, Hayward, CA) at 40°C for 30 min. Then each sample was hybridized with one BaseScope probe according to the requirement in HybEZ™ Oven at 40°C for 2 hours. After buffer washing steps, the samples were incubated in HybEZ™ Oven at 40°C with the serial application of BaseScope™ v2 Amp 1~8 for signal amplification. At last, Fast Red substrate was added to samples for 10 min at RT to detect target RNA and the slides were counterstained with 50% hematoxylin staining solution for 2 min at RT. Target RNA was visualized using a standard bright field microscope.

Zhao M., Ren K., Xiong X., Xin Y., Zou Y., Maynard J.C., Kim A., Battist A.P., Koneripalli N., Wang Y., Chen Q., Xin R., Yang C., Huang R., Yu J., Huang Z., Zhang Z., Wang H., Wang D., Xiao Y., Salgado O.C., Jarjour N.N., Hogquist K.A., Revelo X.S., Burlingame A.L., Gao X., von Moltke J., Lin Z, & Ruan H.B. (2022). Epithelial STAT6 O-GlcNAcylation drives a concerted anti-helminth alarmin response dependent on tuft cell hyperplasia and Gasdermin C. Immunity, 55(4), 623-638.e5.

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RNAscope-Based Adgb mRNA Detection

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RNAscope in situ hybridization was performed using BaseScope Detection Reagent Kit v2 RED (Advanced Cell Diagnostics Inc, Newark, CA, USA, Cat. No. 323900) according to the manufacturer’s instructions. H₂O₂ treatment, antigen retrieval, and protease treatment were performed on 5 µm-thick sections prior to hybridization with probes for Adgb (BA-Mm-Adgb-3zz-st, Advanced Cell Diagnostics, Cat. No. 862141), DapB as negative control (BA-DapB-3zz, Advances Cell Diagnostics, Cat. No. 701011), and Ppib as positive control (Ba-Mm-Ppib-3zz, Advanced Cell Diagnostics, Cat. No. 701071) at 40°C for 2 hr followed by eight amplification steps. The signal was revealed with Fast Red, and the sections were counterstained with Gill’s hematoxylin no. 1 and mounted with VectaMount (Vector Laboratories, Burlingame, CA, USA). The sections were visualized using a light microscope.

Keppner A., Correia M., Santambrogio S., Koay T.W., Maric D., Osterhof C., Winter D.V., Clerc A., Stumpe M., Chalmel F., Dewilde S., Odermatt A., Kressler D., Hankeln T., Wenger R.H, & Hoogewijs D. (2022). Androglobin, a chimeric mammalian globin, is required for male fertility. eLife, 11, e72374.

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Insulin Receptor mRNA Visualization

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To measure whether insulin receptors were selectively depleted in sensory tissue, BaseScope (Advanced Cell Diagnostics) mRNA visualization was used. Tissues were fixed in 10% neutral buffered formalin overnight at room temperature. BaseScope assays were performed on paraffin sections (5 μm thickness Superfrost plus slides, Thermo Fisher Scientific) using guidelines provided by the supplier (Advanced Cell Diagnostics). Briefly, slides were baked at 60°C for 1 hour before deparaffinizing in xylene (2 × 5 min) and ethanol (2 × 2 min); they were then dried at 60°C for 5 minutes. RNAScope hydrogen peroxide was applied for 10 minutes at room temperature, then target retrieval was applied for 15 minutes at 100°C. RNAScope protease III was then applied for 30 minutes at 40°C. BaseScope insulin receptor (catalog 719141), positive control (catalog 01071), and negative control probes (catalog 701011) were purchased from Advanced Cell Diagnostics and in situ hybridized to tissue sections with BaseScope Detection Reagent Kit v2 - RED (Advanced Cell Diagnostics). Slides were counterstained with 50% Gill’s hematoxylin and then 0.02% ammonia water before drying for 15 minutes at 60°C and mounting in EcoMount (Biocare Medical).

Calco G.N., Maung J.N., Jacoby D.B., Fryer A.D, & Nie Z. (2022). Insulin increases sensory nerve density and reflex bronchoconstriction in obese mice. JCI Insight, 7(20), e161898.

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In Situ Hybridization of Molecular Markers

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Tissue sections were air-dried at 60 °C for 1 hr to overnight. RNAScope multiplex fluorescent reagent kit v2 (323100, Advanced Cell Diagnostics) and BaseScope detection reagent kit v2-Red (323910, Advanced Cell Diagnostics) were used for in situ hybridization according to the manufacturer’s instructions. RNAScope probes from Advanced Cell Diagnostics used in this study were Aldh1a2 (447391), Tbx15 (558761), Smoc2 (318514), Fgf18 (495421), Fgfr1 (443491), Fgfr2 (443501 s), Fgfr3 (440771), Fgfr4 (443511), Lama4 (494901), Hic1 (464131), Creb5 (572891), Meox1 (530641-C2), Etv5 (316961), Bach2 (887121-C3), Myod1 (316081 or 316081-C2), Myf5 (492911) and Fgf18-Exon1C (1118021-C1).

Feng J., Han X., Yuan Y., Cho C.K., Janečková E., Guo T., Pareek S., Rahman M.S., Zheng B., Bi J., Jing J., Zhang M., Xu J., Ho T.V, & Chai Y. (2022). TGF-β signaling and Creb5 cooperatively regulate Fgf18 to control pharyngeal muscle development. eLife, 11, e80405.

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In Situ Hybridization of Mttp RNA in Mouse Retina

Cited 2 times

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Mttp RNA in situ hybridization was performed on fixed frozen sections using the BaseScope 2.5 HD assay kit (Advanced Cell Diagnostics, Newark, CA) (32 (link), 33 (link)). A custom-designed BaseScope probe targeting bases 855–989 of the mouse Mttp transcript, NM_008642.3, which span exon 5 and part of exon 6 was used for the assay (BaseScope^™ Probe- BA-Mm-Mttp-3zz-st-C1; Cat # 1217461-C1; Advanced Cell Diagnostics, Minneapolis, Minnesota, USA). Briefly, mouse retinal cryosections (10 μm thick) from ~12 month-old RPEΔMttp and control mice retinas were post-fixed in 4% paraformaldehyde (PFA) for 60 mins at 4°C, followed by dehydration in ethanol, bleaching (10% H2O2 at 60°C), target retrieval (5 min), dehydration, and Protease III treatment (15 min at 40°C). The sections were incubated with the BaseScope probe for 2 h at 40°C, followed by Hybridize BaseScope^™ v2 AMP steps 1–8 and signal detection per manufacturer’s instructions (BaseScope^™ Detection Reagent Kit v2 – RED; Document # 323900-USM; Advanced Cell Diagnostics). The sections were counterstained with Hoechst 33258 (1:10,000) and mounted with Prolong Gold Antifade Mountant (P36930, Molecular Probes, Waltham, MA, USA).

Grubaugh C.R., Dhingra A., Prakash B., Montenegro D., Sparrow J.R., Daniele L.L., Curcio C.A., Bell B.A., Hussain M.M, & Boesze-Battaglia K. (2023). Microsomal triglyceride transfer protein is necessary to maintain lipid homeostasis and retinal function. bioRxiv.

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