CUBIC-HV™1 3D nuclear staining kit

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Einheit	Stückpreis	Belgien	Japan*
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*Lagerbestand in Belgien: Versand am selben Tag
*Lagerbestand in Japan: Bitte prüfen Sie die Versandsimulation für voraussichtliche Lieferzeiten. (ausgenommen regulierte Produkte und Trockeneissendungen)

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This kit contains buffers and typical nuclear stains for 3D nuclear staining of CUBIC-cleared tissues.

Please see the detail protocols in CUBICStars’ web site.
https://www.cubicstars.com/cubic-hv/index.html

Constituents in kit (suitable for a brain of an adult mouse)

• CUBIC-HV™1 3D nuclear staining buffer (1x) (for 10 tests)
• CUBIC-HV™1 3D nuclear staining washing buffer (100x) (for 10 tests)
• DAPI·2HCl (200x) (for 10 tests)
• Propidium Iodide (100x) (for 10 tests)
• 10 x 5 mL tube

Process outline

====Delipidation====

Perfusion fixation and dissection of the mouse brain
Post-fix <24 h
Wash (PBS) 2 h x3
Delipidation (CUBIC-L; Product No. T3740) 3-7 days
Wash (PBS) 2 h x3

====Nuclear staining by CUBIC-HV™1 3D nuclear staining kit (Product No. C3709) ====

Nuclear staining (DAPI or Propidium Iodide or others in CUBIC-HV™1 3D nuclear staining buffer) 3-5 days
Wash (3D nuclear staining wash buffer; 10mM HEPES buffer, pH 7.5) 2 h x3

==== Optional process; Enzyme reaction ====

====Immunostaining by CUBIC-HV™1 3D immunostaining kit (Product No. C3708) ====

Primary antibody (anti-NeuN antibody in C3708 kit can be used): 5 µg/whole brain
Secondary antibody: anti-Fc Fab (out of the kits), 3.75 µg/whole brain
Primary antibody + secondary antibody reaction / Replacement with CUBIC-HV™1 3D immunostaining buffer, 1.5 h
Immunostaining (CUBIC-HV™1 3D immunostaining buffer with CUBIC-HV™1 3D immunostaining additive (if required)), >1 week
Reaction in the same buffer at 4°C, 1 day
Wash (CUBIC-HV™1 3D immunostaining wash buffer), 2 h x3

====Fix and RI matching====

Post-fix (in 1% Formaldehyde Solution), 1 day
Wash (PBS) 2 h
RI matching (CUBIC-R+; Product No. T3740), <3 days
Microscopic observation

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