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NACRES:
NA.25
UNSPSC Code:
12161700
Servicio técnico
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Molecular Biology
Quality Level
form
lyophilized powder
technique(s)
Southern blotting: suitable
foreign activity
DNAses, RNases, NICKases, endonucleases, and exonucleases, none detected
storage temp.
2-8°C
General description
Denhardt′s Solution is a blocking agent routinely used to reduce non-specific binding of detection reagents to the Northern and Southern Blot membranes. This powder can be used to make a 50x concentrate that can be diluted to any concentration for use in a variety of pre-hybridization and hybridization buffers.
Application
Suitable as a blocking Buffer for Southern and Northern blot membranes.
Denhardt′s Solution has been used:
- as a component in hybridization buffer for the hybridization of brain sections
- in in-situ hybridization peroxidase-(antiperoxidase method-whole mounts) (ISHPAP-WM) process
- in the preparation of hybridization mixture
Biochem/physiol Actions
Denhardt′s solution and gelatin-chrome alum can be used to treat chromosomes for G-banding with trypsin after in situ hybridization.
Preparation Note
Each vial of Denhardt′s powder contains 50mg Bovine Serum Albumin (BSA), 50mg Ficoll, and 50mg Polyvinylpyrrolidone (PVP).
To make a 50x concentrate, dissolve entire contents of vial in 5 mL molecular biology grade water. Store at -20°C for up to two years.
Clase de almacenamiento
11 - Combustible Solids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, type N95 (US)
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Sambrook, J., et al.
Molecular Cloning: A Laboratory Manual, 9-9 (1989)
E J Mackie et al.
Journal of cell science, 103 ( Pt 3), 765-771 (1992-11-01)
The extracellular matrix glycoprotein, tenascin, is associated in vivo with mesenchyme undergoing osteogenesis and chondrogenesis, but is absent from mature bone and cartilage matrix. The expression of tenascin by osteoblastic cells in vitro has been investigated by immunoblotting and immunocytochemistry.
M Simpson-Holley et al.
Journal of virology, 85(4), 1581-1593 (2010-12-15)
Vaccinia virus (VV) mutants lacking the double-stranded RNA (dsRNA)-binding E3L protein (ΔE3L mutant VV) show restricted replication in most cell types, as dsRNA produced by VV activates protein kinase R (PKR), leading to eIF2α phosphorylation and impaired translation initiation. Here