Home › Immunology › Immunohistochemical Detection of 5-Methylcytosine and 5-Hydroxymethylcytosine in Developing and Postmitotic Mouse Retina
Immunology JoVE (Open Access) Citable · DOI

Immunohistochemical Detection of 5-Methylcytosine and 5-Hydroxymethylcytosine in Developing and Postmitotic Mouse Retina

DOI: 10.3791/58274-v
What you'll learn
  • ✓ Prepare mouse retinal tissue for immunohistochemical staining
  • ✓ Perform immunostaining with 5mC and 5hmC antibodies
  • ✓ Interpret epigenetic marker distribution in developing retina
Protocol

Biopharma Insights The objective of this report is to describe the protocol for robust immunohistochemical detection of epigenetic markers, 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) in developing and postmitotic mouse retina.

Difficulty
intermediate
Total time
~4–6 hours per sample (excluding tissue collection)
Model organism
Mouse retina (developmental and postmitotic)
Biosafety
BSL-1

Steps

1
Prepare retinal tissue for immunostaining

Collect mouse retinal tissue and prepare it for immunohistochemical analysis. This includes fixation, sectioning, and mounting on microscope slides.

▶ 00:59
2
Perform immunostaining with 5mC and 5hmC antibodies

Apply primary antibodies against 5-methylcytosine and 5-hydroxymethylcytosine to tissue sections, followed by secondary antibody detection to visualize epigenetic modifications.

▶ 03:02
3
Analyze marker distribution in retinal development

Examine and document immunostaining patterns of 5mC and 5hmC across developing and mature retinal tissue to characterize epigenetic changes.

▶ 05:02
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