In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
BIX-01294 is a small molecule inhibitor of G9a and GLP (a H3K9 PKMT that shares 80% sequence identity with G9a in their respective SET domains) that was discovered via screening of a library of 125,000 synthetic compounds. BIX-01294 is selective for G9a and GLP over several H3K9 PKMTs including SUV39H1 and ESET, other KMTs such as SET7/9, and the arginine methyltransferase PRTM1. The X-ray crystal structure of GLP and BIX-01294 confirmed that BIX-01294 bound to the histone peptide binding pocket but failed to interact with the lysine binding channel. Cells dosed with BIX-01294 at 4.1 μM were characterized by reduced H3K9me2 levels in several cell lines but toxicity to cells at > 4.1 μM was observed.
William P. Janzen, Tim J. Wigle, Jian Jin, Stephen V. Frye. Epigenetics: tools and technologies. Drug Discov Today Technol. 2010 ; 7(1): 59 – 65.
|Physical Appearance||A solid|
|Storage||Store at -20°C|
|Solubility||≥24.45mg/mL in DMSO|
|Shipping Condition||Evaluation sample solution : ship with blue ice.All other available size: ship with RT , or blue ice upon request|
|General tips||For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.|
|Kinase experiment :|
Dissociation Enhanced Lanthanide Fluoro-Immuno Assay (DELFIA) was performed in white, opaque 384-well plates coated with Neutravidin. Test compounds were diluted to 12 μg/mL in 50mM Tris-HCl (pH 8.5) containing 4% DMSO and 10 μL was dispensed into the wells. Blank and control wells received only compound buffer. GST-G9a at 10 μg/mL and SAM at 40 μM were diluted in 50mM Tris HCl (pH 8.5)/10 mM DTT and added in a volume of 20 μL. Blank wells received Tris/DTT buffer only. The reactions were initiated by the addition of 800 nM H3(1-20)-cysbiotin substrate in 50 mM Tris (pH 8.5) in a volume of 10 μL, and incubated at room temperature for 60 mins. The plates were washed 3 times with 100 μL of Wash Buffer. Next, 50 μL of Fluoroimmunoassay (FI) Buffer containing 5 ng α-2X-di-meth H3-K9 and 5 ng goat anti-rabbit Eu chelate was added to all wells of the plate, and the plate was incubated for an additional 1 hr at room temperature. The plates were washed 3 times with 100 μL of Wash Buffer, and 50 μL of Enhancement Solution was added to each well. Time resolved fluorescence was measured after 45 mins on a Viewlux Microplate Imager imaging for 15 seconds.
|Cell experiment :|
Wild-type and G9a deficient mouse embryonic stem (ES) cells, mouse embryonic fibroblasts and human HeLa cells
The solubility of this compound in DMSO is > 10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.
4.1 μM; over 2 days
BIX-01294 (4.1 μM) reduced the H3K9me2 Levels in Bulk Histone Preparations from wild-type mouse ES cells, mouse embryonic fibroblasts and human HeLa cells, but not in G9a deficient mouse ES cells.
. Kubicek S, O'Sullivan RJ, August EM, Hickey ER, Zhang Q, Teodoro ML, Rea S, Mechtler K, Kowalski JA, Homon CA, Kelly TA, Jenuwein T. Reversal of H3K9me2 by a small-molecule inhibitor for the G9a histone methyltransferase. Mol Cell. 2007 Feb 9;25(3):473-81.
|Description||BIX 01294 is an inhibitor of G9a-like protein and G9a histone lysine methyltransferase with IC50 value of 0.7 and 1.7 μM, respectively.|
|Targets||G9a-like protein||G9a histone lysine methyltransferase|
|IC50||0.7 μM||1.7 μM|