Key Spec Table
|Species Reactivity||Key Applications||Host||Format||Antibody Type|
|H||WB, ICC, IP||M||Purified||Monoclonal Antibody|
|Presentation||Purified mouse monoclonal IgG2b in buffer containing 0.1 M Tris-Glycine, (pH 7.4), 150 mM NaCl with 0.05% sodium azide.|
|Application||Anti-Mad1 Antibody, clone 9B10 is an antibody against Mad1 for use in WB, IC & IP.|
|Safety Information according to GHS|
|Storage and Shipping Information|
|Storage Conditions||Stable for 1 year at 2-8ºC from date of receipt.|
|Material Size||100 µg|
|Anti-Mad1, clone 9B10 - 2531919||2531919|
|Anti-Mad1, clone 9B10 - NG1578247||NG1578247|
|Anti-Mad1, clone 9B10_2842489||2842489|
|Reference overview||Species||Pub Med ID|
|Sulforaphane causes epigenetic repression of hTERT expression in human breast cancer cell lines.|
Meeran, SM; Patel, SN; Tollefsbol, TO
PloS one 5 e11457 2010
Sulforaphane (SFN), an isothiocyanate found in cruciferous vegetables, is a common dietary component that has histone deacetylase inhibition activity and exciting potential in cancer prevention. The mechanisms by which SFN imparts its chemopreventive properties are of considerable interest and little is known of its preventive potential for breast cancer.We found that SFN significantly inhibits the viability and proliferation of breast cancer cells in vitro while it has negligible effects on normal breast cells. Inhibition of telomerase has received considerable attention because of its high expression in cancer cells and extremely low level of expression in normal cells. SFN treatment dose- and time-dependently inhibited human telomerase reverse transcriptase (hTERT), the catalytic regulatory subunit of telomerase, in both MCF-7 and MDA-MB-231 human breast cancer cells. DNA methyltransferases (DNMTs), especially DNMT1 and DNMT3a, were also decreased in SFN-treated breast cancer cells suggesting that SFN may repress hTERT by impacting epigenetic pathways. Down-regulation of DNMTs in response to SFN induced site-specific CpG demethylation occurring primarily in the first exon of the hTERT gene thereby facilitating CTCF binding associated with hTERT repression. Chromatin immunoprecipitation (ChIP) analysis of the hTERT promoter revealed that SFN increased the level of active chromatin markers acetyl-H3, acetyl-H3K9 and acetyl-H4, whereas the trimethyl-H3K9 and trimethyl-H3K27 inactive chromatin markers were decreased in a dose-dependent manner. SFN-induced hyperacetylation facilitated the binding of many hTERT repressor proteins such as MAD1 and CTCF to the hTERT regulatory region. Depletion of CTCF using siRNA reduced the SFN-induced down-regulation of hTERT mRNA transcription in these breast cancer cells. In addition, down-regulation of hTERT expression facilitated the induction of cellular apoptosis in human breast cancer cells.Collectively, our results provide novel insights into SFN-mediated epigenetic down-regulation of telomerase in breast cancer prevention and may open new avenues for approaches to SFN-mediated cancer prevention.Full Text Article
|Tripin/hSgo2 recruits MCAK to the inner centromere to correct defective kinetochore attachments.|
Huang, Haomin, et al.
J. Cell Biol., 177: 413-24 (2007) 2007
|Mapping the assembly pathways that specify formation of the trilaminar kinetochore plates in human cells.|
Liu, Song-Tao, et al.
J. Cell Biol., 175: 41-53 (2006) 2006
We report the interactions amongst 20 proteins that specify their assembly to the centromere-kinetochore complex in human cells. Centromere protein (CENP)-A is at the top of a hierarchy that directs three major pathways, which are specified by CENP-C, -I, and Aurora B. Each pathway consists of branches that intersect to form nodes that may coordinate the assembly process. Complementary EM studies found that the formation of kinetochore trilaminar plates depends on the CENP-I/NUF2 branch, whereas CENP-C and Aurora B affect the size, shape, and structural integrity of the plates. We found that hMis12 is not constitutively localized at kinetochores, and that it is not essential for recruiting CENP-I. Our studies also revealed that kinetochores in HeLa cells contain an excess of CENP-A, of which approximately 10% is sufficient to promote the assembly of normal levels of kinetochore proteins. We elaborate on a previous model that suggested kinetochores are assembled from repetitive modules (Zinkowski, R.P., J. Meyne, and B.R. Brinkley. 1991. J. Cell Biol. 113:1091-110).