PMID- 33326080
OWN - NLM
STAT- MEDLINE
DCOM- 20210329
LR  - 20210329
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2254
DP  - 2021
TI  - A Quick Immuno-FISH Protocol for Detecting RNAs, Proteins, and Chromatin 
      Modifications.
PG  - 251-257
LID - 10.1007/978-1-0716-1158-6_15 [doi]
AB  - Immunofluorescence and fluorescence in situ hybridization (FISH) are widely used 
      cytogenetic techniques for visualization of protein and RNA/DNA molecules. Here, 
      we describe an experimental procedure for quick sequential immunofluorescence and 
      RNA FISH (immuno-FISH), which enables the simultaneous detection of proteins, 
      chromatin modifications, and RNAs on the inactive X-chromosome (Xi) using female 
      mouse embryonic fibroblast (MEF) and tail-tip 3T3 cell lines. Using a pooled 
      array of oligonucleotides labeled with a single fluorophore as an RNA FISH probe, 
      we can reduce the time for RNA FISH from an overnight process to 1-2 h without 
      losing its sensitivity. This protocol could be applied to visualization of 
      various protein and RNA molecules, and chromatin modifications.
FAU - Ogawa, Akiyo
AU  - Ogawa A
AD  - Division of Reproductive Sciences, Cincinnati Children's Hospital Medical Center, 
      Cincinnati, OH, USA.
FAU - Ogawa, Yuya
AU  - Ogawa Y
AD  - Division of Reproductive Sciences, Cincinnati Children's Hospital Medical Center, 
      Cincinnati, OH, USA. yuya.ogawa@cchmc.org.
AD  - Department of Pediatrics, University of Cincinnati College of Medicine, 
      Cincinnati, OH, USA. yuya.ogawa@cchmc.org.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Chromatin)
RN  - 0 (Histones)
RN  - 0 (RNA, Long Noncoding)
RN  - 0 (XIST non-coding RNA)
RN  - 63231-63-0 (RNA)
SB  - IM
MH  - Animals
MH  - Cells, Cultured
MH  - Chromatin/*genetics
MH  - Chromatin Assembly and Disassembly
MH  - Female
MH  - Fluorescent Antibody Technique
MH  - Histones/*metabolism
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Mice
MH  - NIH 3T3 Cells
MH  - RNA/*genetics
MH  - RNA, Long Noncoding/*genetics
MH  - X Chromosome/genetics
OTO - NOTNLM
OT  - Chromatin modifications
OT  - Fluorescent in situ hybridization (FISH)
OT  - Immunofluorescence
OT  - Long noncoding RNA (lncRNA)
OT  - X-chromosome inactivation
OT  - Xist RNA
EDAT- 2020/12/17 06:00
MHDA- 2021/03/30 06:00
CRDT- 2020/12/16 12:13
PHST- 2020/12/16 12:13 [entrez]
PHST- 2020/12/17 06:00 [pubmed]
PHST- 2021/03/30 06:00 [medline]
AID - 10.1007/978-1-0716-1158-6_15 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2021;2254:251-257. doi: 10.1007/978-1-0716-1158-6_15.