PMID- 19462231
OWN - NLM
STAT- MEDLINE
DCOM- 20091125
LR  - 20211020
IS  - 1573-6903 (Electronic)
IS  - 0364-3190 (Linking)
VI  - 34
IP  - 11
DP  - 2009 Nov
TI  - An improved method to obtain a soluble nuclear fraction from embryonic brain 
      tissue.
PG  - 2022-9
LID - 10.1007/s11064-009-9993-9 [doi]
AB  - This paper describes modifications of the standard methods for obtaining a 
      soluble nuclear fraction from embryonic brain tissue. The main improvements are: 
      (1) the inclusion of a low speed centrifugation step to prevent the appearance of 
      high density contaminants, (2) a sucrose density gradient to remove perinuclear 
      mitochondria and ER membranes and (3) a protein extraction approach which 
      significantly enhances protein yield. To demonstrate the effectiveness of the 
      method, pellets were analyzed by light and electron microscopy and purity of the 
      soluble extracts was immunologically tested. Finally, to illustrate the 
      applicability of this approach, the induction of the transcription factor HIF-1 
      (hypoxia-inducible factor-1) was assessed by Western blot using soluble nuclear 
      fractions and by immuno-electron microscopy using purified nuclear fractions, 
      both obtained from the optic lobes of chick embryos. In conclusion, the procedure 
      presently described appears to be reliable and convenient for obtaining a pure 
      soluble nuclear fraction from a discrete amount of embryonic brain tissue.
FAU - Giusti, Sebastian
AU  - Giusti S
AD  - Institute of Cell Biology and Neuroscience "Prof. E De Robertis", University of 
      Buenos Aires, Paraguay, Argentina.
FAU - Bogetti, Maria Eugenia
AU  - Bogetti ME
FAU - Bonafina, Antonela
AU  - Bonafina A
FAU - Fiszer de Plazas, Sara
AU  - Fiszer de Plazas S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090522
PL  - United States
TA  - Neurochem Res
JT  - Neurochemical research
JID - 7613461
RN  - 0 (Hypoxia-Inducible Factor 1)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Surface-Active Agents)
RN  - 368GB5141J (Sodium Dodecyl Sulfate)
RN  - 9002-93-1 (Octoxynol)
SB  - IM
MH  - Animals
MH  - Brain/*ultrastructure
MH  - Cell Fractionation/*methods
MH  - *Cell Nucleus/chemistry/ultrastructure
MH  - Centrifugation
MH  - Chick Embryo
MH  - Hypoxia-Inducible Factor 1/biosynthesis
MH  - Microscopy, Immunoelectron
MH  - Nuclear Proteins/isolation & purification
MH  - Octoxynol
MH  - Sodium Dodecyl Sulfate
MH  - Solubility
MH  - Surface-Active Agents
EDAT- 2009/05/23 09:00
MHDA- 2009/12/16 06:00
CRDT- 2009/05/23 09:00
PHST- 2009/03/25 00:00 [received]
PHST- 2009/05/07 00:00 [accepted]
PHST- 2009/05/23 09:00 [entrez]
PHST- 2009/05/23 09:00 [pubmed]
PHST- 2009/12/16 06:00 [medline]
AID - 10.1007/s11064-009-9993-9 [doi]
PST - ppublish
SO  - Neurochem Res. 2009 Nov;34(11):2022-9. doi: 10.1007/s11064-009-9993-9. Epub 2009 
      May 22.