Simple or double in situ hybridization and immunofluorescent staining were performed on transverse sections as previously reported (Touahri et al., 2012 ; Ventéo et al., 2012 ). Digoxigenin‐ or Fluorescein‐labeled antisens RNA probes for sulf2, aldh1L1, fgfr3, sox10, and tenascin‐C were synthesized using DIG‐ or Fluorescein‐labelling kit (Roche) according to the manufacturer's instructions and revealed using either NBT/BCIP (Roche), INT/BCIP (Roche) or Alexa‐fluor 488 Tyramide (Molecular Probes). Antibodies used in this study were as follows: goat anti‐AldoC (Santa Cruz Biotechnology), rabbit and mouse anti‐GFAP (DAKO), rabbit anti‐NFIA (Active Motif), rabbit and goat anti‐Olig2 (Millipore and R&D Systems), goat anti‐Olig1 (R&D Systems), goat anti‐Sox10 (Santa Cruz Biotechnology), rat anti‐PDGFRα (BD Pharmingen), rabbit anti‐Zeb1 (Novus), mouse anti‐Nkx6.1 (Hybridoma Bank), mouse anti‐Cx43 (BD Biosciences) and rabbit anti‐Cx30 (Thermo Fisher Scientific). Alexa Fluor‐594‐ or Alexa Fluor‐488 or Alexa Fluor‐647‐conjugated secondary antibodies (Thermo Fisher Scientific) were used.
Dig or fluorescein labelling kit
Manufactured by Roche
The DIG- or Fluorescein-labelling kit is a laboratory tool used for the labelling of biomolecules, such as nucleic acids or proteins, with either digoxigenin (DIG) or fluorescein compounds. The kit provides the necessary reagents and protocols to enable the attachment of these labels to the target molecules, facilitating their detection and analysis in various downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
Nbt bcip, by Roche (2 mentions)
Goat anti sox10, by Santa Cruz Biotechnology (2 mentions)
Tyramide alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Mouse anti cx43, by BD (1 mentions)
Int bcip, by Roche (1 mentions)
Rabbit anti cx30, by Thermo Fisher Scientific (1 mentions)
Rabbit anti nfia, by Active Motif (1 mentions)
Rabbit anti zeb1, by Novus Biologicals (1 mentions)
Rat anti pdgfrα, by BD (1 mentions)
Alexa fluor 647 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Rabbit anti olig2, by Merck Group (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
2 protocols using dig or fluorescein labelling kit
1
Transcription and Protein Expression Analysis
2
In Situ Hybridization and Immunostaining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Simple or double in situ hybridization and immunofluorescent stainings were performed on transverse sections as previously reported (Ohayon et al., 2019) (link). Digoxigenin-or Fluoresceinlabeled antisens RNA probes for inka2, efnb3, fgfr3 and kcnip3 were synthesized using DIGor Fluorescein-labelling kit (Roche) according to the manufacturer's instructions and revealed using either NBT/BCIP (Roche), or Alexa-fluor 488 and 555 Tyramide (Thermo Fisher Scientific). Antibodies used in this study were as follows: rabbit anti-Olig2 (Millipore, CatN#AB9610) and goat anti-Sox10 (Santa Cruz Biotech nology, CatN#sc-17342). Alexa
Fluor-488 or Alexa Fluor-647-conjugated secondary antibodies (Thermo Fisher Scientific)
were used.
Fluor-488 or Alexa Fluor-647-conjugated secondary antibodies (Thermo Fisher Scientific)
were used.
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