Biomedical Engineering Reference
In-Depth Information
14. If you want to confirm that the binding protein to the probe
is actually the transcriptional factor of your interest, add the
antibody (around 0.5 mg) against the transcriptional factor to
the lysates at this step and incubate for ~30 min on ice.
Control IgG should be used as a negative control. The band
shift will be further enhanced by binding of antibody to tran-
scriptional factor-probe complex. Alternatively, shifted band
is reduced or disappeared because of perturbation of interac-
tion between transcriptional factor and probe by antibody.
This assay is called Super Shift Assay.
References
1. The Huntington's Disease Collaborative
Research and Group (1993) A novel gene
containing a trinucleotide repeat that is
expanded and unstable on Huntington's dis-
ease chromosomes. Cell 72:971-983
2. Landles C, Bates GP (2004) Huntingtin and
the molecular pathogenesis of Huntington's
disease. Fourth in molecular medicine review
series. EMBO Rep 5:958-963
3. Li SH, Li XJ (2004) Huntingtin-protein inter-
actions and the pathogenesis of Huntington's
disease. Trends Genet 20:146-154
4. Harjes P, Wanker EE (2003) The hunt for
huntingtin function: interaction partners tell
many different stories. Trends Biochem Sci
28:425-433
5. Luthi-Carter R, Hanson SA, Strand AD,
Bergstrom DA, Chun W, Peters NL, Woods
AM, Chan EY, Kooperberg C, Krainc D,
Young AB, Tapscott SJ, Olson JM (2002)
Dysregulation of gene expression in the R6/2
model of polyglutamine disease: parallel
changes in muscle and brain. Hum Mol Genet
11:1911-1926
6. Chan EY, Luthi-Carter R, Strand A, Solano
SM, Hanson SA, DeJohn MM, Kooperberg
C, Chase KO, DiFiglia M, Young AB, Leavitt
BR, Cha JH, Aronin N, Hayden MR, Olson
JM (2002) Increased huntingtin protein
length reduces the number of polyglutamine-
induced gene expression changes in mouse
models of Huntington's disease. Hum Mol
Genet 11:1939-1951
7. Kotliarova S, Jana NR, Sakamoto N, Kurosawa
M, Miyazaki H, Nekooki M, Doi H, Machida
Y, Wong HK, Suzuki T, Uchikawa C,
Kotliarov Y, Uchida K, Nagao Y, Nagaoka U,
Tamaoka A, Oyanagi K, Oyama F, Nukina N
(2005) Decreased expression of hypotha-
lamic neuropeptides in Huntington disease
transgenic mice with expanded polyglu-
tamine-EGFP
fluorescent
aggregates.
J
Neurochem 93:641-653
8. Sugars KL, Rubinsztein DC (2003)
Transcriptional abnormalities in Huntington
disease. Trends Genet 19:233-238
9. Riley BE, Orr HT (2006) Polyglutamine neu-
rodegenerative diseases and regulation of
transcription: assembling the puzzle. Genes
Dev 20:2183-2192
10. Cha JH (2000) Transcriptional dysregulation
in Huntington's disease. Trends Neurosci
23:387-392
11. Yamanaka T, Miyazaki H, Oyama F, Kurosawa
M, Washizu C, Doi H, Nukina N (2008)
Mutant Huntingtin reduces HSP70 expres-
sion through the sequestration of NF-Y tran-
scription factor. EMBO J 27:827-839
12. Doi H, Mitsui K, Kurosawa M, Machida Y,
Kuroiwa Y, Nukina N (2004) Identification
of ubiquitin-interacting proteins in purified
polyglutamine aggregates. FEBS Lett 571:
171-176
13. Busch A, Engemann S, Lurz R, Okazawa H,
Lehrach H, Wanker EE (2003) Mutant hun-
tingtin promotes the fibrillogenesis of wild-
type huntingtin: a potential mechanism for
loss of huntingtin function in Huntington's
disease. J Biol Chem 278:41452-41461
14. Scherzinger E, Lurz R, Turmaine M,
Mangiarini L, Hollenbach B, Hasenbank R,
Bates GP, Davies SW, Lehrach H, Wanker EE
(1997) Huntingtin-encoded polyglutamine
expansions form amyloid-like protein aggre-
gates in vitro and in vivo. Cell 90:549-558
15. Mosser DD, Theodorakis NG, Morimoto RI
(1988) Coordinate changes in heat shock
element-binding activity and HSP70 gene
transcription rates in human cells. Mol Cell
Biol 8:4736-4744
Search WWH ::
Custom Search