Fresh tissues were collected and fixed by 10% neutral buffered formalin for 16–32 h at room temperature. Samples were embedded in paraffin blocks and cut into 5 μm sections for staining. Freshly cut slides were air-dried overnight at room temperature, then baked for 1 h at 60°C. The RNA-Scope in situ hybridization was performed by using RNA-Scope 2.5 HD Detection Reagent Red Kit (Advanced Cell Diagnostics, Newark, CA) following the manufacturer’s standard protocol.
Rna scope 2.5 hd detection reagent red kit
Manufactured by Advanced Cell Diagnostics
Sourced in United States
The RNA-Scope 2.5 HD Detection Reagent Red Kit is a product designed for in situ hybridization analysis. The kit includes reagents necessary for the detection of target RNA molecules within tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
In situ cell death detection kit, by Roche (1 mentions)
Bz x710, by Keyence (1 mentions)
Ecomount, by Biocare Medical (1 mentions)
Superfrost slide, by Thermo Fisher Scientific (1 mentions)
Rnascope 2.5 hd detection reagent brown kit, by Advanced Cell Diagnostics (1 mentions)
Vectamount permanent mounting medium, by Vector Laboratories (1 mentions)
Image pro plus version 6, by Media Cybernetics (1 mentions)
11 protocols using rna scope 2.5 hd detection reagent red kit
1
RNA-Scope in situ Hybridization Protocol
2
Histological Analysis of Pth1r Mutant Bones
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Femura and tibiae were harvested from F1 H223R‐PTH1R/wt‐Pth1r males and females on postnatal days 6, 12, and 27, along with samples from their wt‐Pth1r/wt‐Pth1r littermates. After fixation with 10% formalin for 2 days, bones were rinsed with PBS before being transferred to 70% EtOH. Sections (5 μm) were stained with hematoxylin and eosin (H&E) and analyzed by light microscopy (Keyence BZ‐X710). Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was performed on paraffin sections using an in situ Cell Death Detection kit (Roche, Mannheim, Germany; Catalog No. 11684795910), as previously reported.(31 ) RNA in situ hybridization analysis was carried out on adjacent sections using the RNAscope 2.5 HD Detection Reagent ‐ RED kit (Advanced Cell Diagnostics, Hayward, CA; Catalog No. 322360); the same company provided the probes specific for mRNAs derived from Col2a1 (Catalog No. 407221), Col10a1 (Catalog No. 426181) or PTH1R (Catalog No. 537131).(32 )
3
RNAScope ISH for ASFV Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the in situ hybridization (ISH) assay, the RNAScope 2.5 HD Detection Reagent-Red kit (Advanced Cell Diagnostics, Inc., CA, United States) was used. The procedure was performed following the manufacturer’s instructions. Briefly, tissue sections were cleared and hydrated in xylene and 100% ethanol and then air-dried. The sections were quenched for 10 min in aqueous H2O2, boiled in target retrieval solution for 15 min, rinsed in 100% ethanol and air-dried again. Then a final pre-treatment of protease plus enzyme for 15 min at 40°C was applied. The V-AFSV-01 probe (Advanced Cell Diagnostics, Inc., CA, United States) was applied and incubated at 40°C for 2 h. Sections were then washed twice in 1× wash buffer. After each of the subsequent hybridization steps, the sections were washed twice again. The signal was visualized with the chromogen Fast Red. The sections were then counterstained with Gill’s 1 hematoxylin, dried, cover-slipped with EcoMount (BioCare Medical, CA, United States), and examined by the pathologist. A negative control lymph node from a non-infected animal was tested in an ISH assay, and no immunostaining was observed. In addition, a negative control probe on positive tissue (lymph node from an ASFV Georgia 2007/1 infected animal) was tested, and no staining was observed.
4
RNA-Scope in situ Hybridization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh tissues were collected and fixed by 10% neutral buffered formalin for 16–32 h at room temperature. Samples were embedded in paraffin blocks and cut into 5 μm sections for staining. Freshly cut slides were air-dried overnight at room temperature, then baked for 1 h at 60°C. The RNA-Scope in situ hybridization was performed by using RNA-Scope 2.5 HD Detection Reagent Red Kit (Advanced Cell Diagnostics, Newark, CA) following the manufacturer’s standard protocol.
5
Plin2 mRNA Detection in Nestin-GFP Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Twenty five micrometres sagittal brain sections from transcardially perfused NestinGFP mice were cut on a cryostat, mounted on SuperFrost+ slides (10149870, Fisher Scientific) and dried overnight. Hybridization was performed using the RNAscope 2.5 HD Detection Reagent—RED Kit (322360, Advanced Cell Diagnostics) according to the manufacturer’s instructions. Briefly, hybridization was performed for 2 h at 40 °C in a hybridization oven using branched DNA probes cocktail specific for murine Plin2 mRNA (577111, Advanced Cell Diagnostics), followed by two washes in the provided wash buffer. Six amplification steps were performed subsequently at 40 °C with alternating incubations of 30 and 15 min and washes using the provided wash buffer. Sections were stained, using a triton-based protocol, against GFP as described under “Immunohistochemistry”.
6
Detecting Nipah Virus RNA In Situ
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissues fixed in 10% neutral phosphate buffered formalin were routinely processed, paraffin-embedded and sectioned at 5 µm. Tissue sections were processed as per manufacturer’s instructions for RNAscope® 2.5HD Detection Reagent – Red kit using the V-Nipah-StrM.B.-N probe (Advanced Cell Diagnostics Cat No. 439251). The sections were then counter stained with Gill’s hematoxylin, dehydrated, cleared and coverslipped.
7
In Situ Hybridization for Viral Genomic Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For in situ hybridization (ISH) to detect viral genomic material, five μm paraffin-embedded formalin fixed tissue sections were cut, air dried then melted on to charged slides in a 60 °C oven. The slides were then cleared and dehydrated in xylene and 100% ethanol, then air dried. The sections were quenched for ten minutes in aqueous H2O2, boiled in target retrieval solution for fifteen minutes, rinsed in 100% ethanol and air-dried again. Then a final pre-treatment of protease plus enzyme for fifteen minutes at 40 °C was applied. The probe (V-RESTV-NP-C1, Advanced Cell Diagnostics) was applied and incubated at 40 °C for two hours. The hybridization amplification steps (AMP 1-6) were applied to the slides for the recommended times and temperatures as per the manual for the RNAscope® 2.5HD Detection Reagent—Red kit (Advanced Cell Diagnostics). The signal was then visualized by the chromogen Fast Red. The sections were then counter-stained with Gill’s No. 1 hematoxylin, dried and cover-slipped.
8
FFPE Lung Tissue ISH Analysis
9
Quantifying p16Ink4a Expression in Murine Osteocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA in situ hybridization was performed using the RNAScope 2.5 HD detection reagent RED kit from Advanced Cell Diagnostics (Newark, CA, US) following the manufacturer’s instructions, as previously described (64 ). The following probes were incubated on paraffin-embedded tissue sections for 2 hours at 40C: murine p16Ink4a (Cat#411011) and positive/negative controls (Cat#313911, Cat#310043). The signal was detected for 10 min at RT. Sections were counterstained with hematoxylin, dehydrated at 60C for 20 min, and mounted with VectaMount permanent mounting medium (Vector Laboratories, Newark, CA, US). The number of positive/negative osteocytes was quantified using a brightfield microscope at 40X magnification. Analyses were performed in the cortical bone of an 800-μm region of the tibia, starting 200 μm below the growth plate, in a blinded fashion by two independent investigators.
10
RNA-Scope Analysis of Stromal Cells
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!