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A modified immunofluorescence in situ hybridization method to detect long non‑coding RNAs and proteins in frozen spinal cord sections

  • Authors:
    • Chong Meng
    • Xin Zhao
    • Jie Lao
  • View Affiliations / Copyright

    Affiliations: Department of Hand Surgery, Huashan Hospital, Fudan University, Shanghai 200040, P.R. China
    Copyright: © Meng et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 4.0].
  • Pages: 4623-4628
    |
    Published online on: April 10, 2018
       https://doi.org/10.3892/etm.2018.6046
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Abstract

Immunofluorescence in situ hybridization (immuno‑FISH) is widely used to co‑detect RNAs and proteins in order to study their spatial distribution in cells. The present study used a modified immuno‑FISH protocol for the detection of long non‑coding RNAs (lncRNAs) and proteins in frozen spinal cord sections. The spinal cords of Sprague‑Dawley rats were harvested, frozen and sectioned (10 µm), and oligonucleotide probes and antibodies were prepared. Following antigen retrieval, dehydration, prehybridization, hybridization, post‑hybridization and immunofluorescence staining, images were captured. Antigen retrieval was performed by autoclaving or proteinase K treatment, and their effects on the hybridization signal were compared. The same sections were successfully stained by immunofluorescence. Satisfactory fluorescent signals of lncRNA and protein were obtained. The results of the present study suggest that the modified protocol of immuno‑FISH for the detection of lncRNAs and proteins in frozen spinal cord sections is effective and time‑efficient, and the required reagents are readily available.
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Copy and paste a formatted citation
Spandidos Publications style
Meng C, Zhao X and Lao J: A modified immunofluorescence in situ hybridization method to detect long non‑coding RNAs and proteins in frozen spinal cord sections. Exp Ther Med 15: 4623-4628, 2018.
APA
Meng, C., Zhao, X., & Lao, J. (2018). A modified immunofluorescence in situ hybridization method to detect long non‑coding RNAs and proteins in frozen spinal cord sections. Experimental and Therapeutic Medicine, 15, 4623-4628. https://doi.org/10.3892/etm.2018.6046
MLA
Meng, C., Zhao, X., Lao, J."A modified immunofluorescence in situ hybridization method to detect long non‑coding RNAs and proteins in frozen spinal cord sections". Experimental and Therapeutic Medicine 15.6 (2018): 4623-4628.
Chicago
Meng, C., Zhao, X., Lao, J."A modified immunofluorescence in situ hybridization method to detect long non‑coding RNAs and proteins in frozen spinal cord sections". Experimental and Therapeutic Medicine 15, no. 6 (2018): 4623-4628. https://doi.org/10.3892/etm.2018.6046
Copy and paste a formatted citation
x
Spandidos Publications style
Meng C, Zhao X and Lao J: A modified immunofluorescence in situ hybridization method to detect long non‑coding RNAs and proteins in frozen spinal cord sections. Exp Ther Med 15: 4623-4628, 2018.
APA
Meng, C., Zhao, X., & Lao, J. (2018). A modified immunofluorescence in situ hybridization method to detect long non‑coding RNAs and proteins in frozen spinal cord sections. Experimental and Therapeutic Medicine, 15, 4623-4628. https://doi.org/10.3892/etm.2018.6046
MLA
Meng, C., Zhao, X., Lao, J."A modified immunofluorescence in situ hybridization method to detect long non‑coding RNAs and proteins in frozen spinal cord sections". Experimental and Therapeutic Medicine 15.6 (2018): 4623-4628.
Chicago
Meng, C., Zhao, X., Lao, J."A modified immunofluorescence in situ hybridization method to detect long non‑coding RNAs and proteins in frozen spinal cord sections". Experimental and Therapeutic Medicine 15, no. 6 (2018): 4623-4628. https://doi.org/10.3892/etm.2018.6046
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