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.
Splitomicin is a selective inhibitor of Sir2p with IC50 value of 60 μM and also a inhibitor of fMLP-induced free radicals with IC50 value of 40.79 ± 9.85 μM [1, 3].
Inhibition of the HDA of Sir2p was the most likely mechanism by which splitomicin caused its phenotypic changes. The direct target of splitomicin is Sir2p deacetylase activity. In addition, splitomicin specific inhibited fMLP-induced superoxide anion release.
In vitro, splitomicin inhibitd NAD+-dependent histone deacetylase activity of the Sir2 protein with an IC50 value of 60μM. By using a [3H]-acetylated histone H4 peptide and measuring the NAD+- dependent release of free [3H]acetate in the presence of whole yeast cell extract from an hst2 strain overexpressing yeast SIR2, a cell extract was obtained from a SIR2-overexpressing hst2 strain. The result established Sir2p deacetylase activity as a direct target of splitomicin. In addition, neutrophils induced by either fMLP (1 μM) or PMA (100 nM) were observed using a flow cytometer and the intracellular production of superoxide anions was investigated at different splitomicin concentrations. Splitomicin inhibited fMLP-induced Mac-1 expressionand increase cAMP levels in human neutrophils [1, 3].
Splitomicin’s naphthoic moiety might be responsible for its inhibitory effects on platelets. By using washed human platelets, the inhibitory effects of splitomicin on platelet aggregation were studied and platelet aggregation and ATP release induced by thrombin (0.1 U/ml), collagen (2 μg/ml), arachidonic acid (0.5 mM), U46619 (2 μM) or ADP (10 μM) was monitored. Splitomicin inhibited platelet aggregation in a concentration dependent manner. Splitomicin increased cAMP and this effect was enhanced when splitomicin (150 μM) was combined with PGE1 (0.5 μM). The inhibitory mechanism of splitomicin on platelet aggregation may increase cyclic AMP levels via inhibition of cyclic AMP phosphodiesterase activity and subsequent inhibition of intracellular Ca ion mobilization, TXB2 formation and ATP release .
. Bedalov A, Gatbonton T, Irvine WP, et al. Identification of a small molecule inhibitor of Sir2p. Proceedings of the National Academy of Sciences of the United States of America, 2001, 98(26): 15113-15118.
. Liu FC, Liao CH, Chang YW, et al. Splitomicin suppresses human platelet aggregation via inhibition of cyclic AMP phosphodiesterase and intracellular Ca++ release. Thrombosis Research, 2009, 124(2): 199-207.
. Liu FC, Day YJ, Liou JT, et al. Splitomicin inhibits fMLP-induced superoxide anion production in human neutrophils by activate cAMP/PKA signaling inhibition of ERK pathway. European Journal of Pharmacology, 2012, 688: 68-75.
|Physical Appearance||A solid|
|Storage||Store at -20°C|
|Solubility||≥9.05mg/mL in DMSO, ≥44.2 mg/mL in EtOH with ultrasonic, <2.3 mg/mL in H2O|
|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.|