KN-93 Phosphate

Catalog No.S7423

KN-93 Phosphate Chemical Structure

Molecular Weight(MW): 599.03

KN-93 Phosphate is a potent and specific inhibitor of Ca2+/calmodulin-dependent protein kinase II (CaMKII) with Ki of 0.37 μM, no remarkable inhibitory effects on APK, PKC, MLCK or Ca2+-PDE activities.

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Cited by 3 Publications

1 Customer Review

  • Paeoniflorin (PF) protects the heart against MI-induced injury in DM mice. Heart function was analyzed by measuring fractional shortening {FS = [LV end diastolic diameter (LVEDD)-LVend systolic diameter (LVESD)]×100/LVEDD} and LV ejection fraction [LVEF = (LVEDD2−LVESD2)/LVEDD2].

    Cell Biosci, 2016, 6:37.. KN-93 Phosphate purchased from Selleck.

Purity & Quality Control

Choose Selective CaMK Inhibitors

Biological Activity

Description KN-93 Phosphate is a potent and specific inhibitor of Ca2+/calmodulin-dependent protein kinase II (CaMKII) with Ki of 0.37 μM, no remarkable inhibitory effects on APK, PKC, MLCK or Ca2+-PDE activities.
Targets
CaMKII [1]
(Cell-free assay)
0.37 μM(Ki)
In vitro

KN-93 inhibits dopamine formation in PC12h cells by modulating the reaction rate of TH to reduce the Ca(2+)-mediated phosphorylation levels of the TH molecule. [1] KN-93 inhibits serum-induced fibroblast cell growth with IC50 of 8 μM, and induces apoptosis after prolonged G1 arrest. [2] KN-93 inhibits androgen receptor activity and p53-independently induces cell death in PCa cells. [3]

Assay
Methods Test Index PMID
Western blot
p-CDK2; 

PubMed: 21448926     


HeLa cells were treated with 100 nM KN-93 or negative control, KN-92. Samples were collected at one hour and four hours post-treatment and cell lysates analyzed by immunoblots for level of Cdk2 T-loop phosphorylation. Inhibition of CamK by KN-93 reduces the level of phosphorylated Cdk2 compared to control KN-92 treated cells.

p-CDK9 / CDK9 ; 

PubMed: 21448926     


HeLa cells were treated with 100 nM KN-93 (CaMK inhibitor) or KN-92 (negative control). Cell lysates prepared after one hour and four hours of treatment were analyzed in an immunoblot for effects on Cdk9 T-loop phosphorylation. After normalization to the Cdk7 loading control and compared to the negative control KN-92, quantification of the immunoblot showed a ~ 55% and ~59% reduction of Cdk9 T-loop phosphorylation at one and four hours, respectively, when CaMK was inhibited by KN-93. Total Cdk9 levels were reduced ~15% at one and four hours by treatment with KN-93.

21448926
In vivo KN-93 (5 μg) ameliorates levodopa-induced dyskinesia by lowering the expression of pGluR1S845 in a rat model of Parkinson’s disease. [4] In MRL/lpr Foxp3-GFP mice, KN-93 results in a significant induction of Treg cells in the spleen, peripheral lymph nodes and peripheral blood, and decreases skin and kidney damage. [5]

Protocol

Kinase Assay:[1]
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Measurement of activities of autophosphorylated/non-autophosphorylate CaMKII:

CaMKII activity is measured utilizing syntideII as a substrate. Purified CaMKII is pre-incubated in the assay mixture ( 35 mM Hepes-Na ( pH 8.0 ), 10 mM MgC12, 0.5 μM CaM, 5 μM ATP, 1 mM CaCl2 or 1 mM EGTA, total 25 μL) at 30 °C for 2 minutes. After this pre-incubation, the protein substrate/radioactive ATP mixture is added to the same test tube and the preparation is further incubated at 30 °C, for 5 minutes ( final assay condition; 35 mM Hepes-Na (pH 8.0), 10 mM MgCl2, 0.125 μM CaCl2, 20 μM syntideII, 11.25 μM [ γ-32P] ATP, 10 % DMSO and indicated concentrations of KN-93, supplemented with 0.25 mM CaCl2 and 2 mM EGTA (for autophosphorylated samples) or 0.25 mM EGTA and 2 mM CaCl2 (for nonautophosphorylated samples ), total 100 μL ). The reaction is terminated by adding of 25 μL of 100 % ( w/v ) ice-cold TCA. After centrifugation, 80 μL of the supernatant is applied to phosphocellulose paper. The filters are then washed with 75 mM H3P04 for 15 min with continuous agitation. After 4-cycles of washing, the radioactivity retains on the filter paper is quantified in a liquid scintillation counter.
Cell Research:[2]
+ Expand
  • Cell lines: NIH 3T3 fibroblasts
  • Concentrations: ~24 μM
  • Incubation Time: 70 hours
  • Method: NIH 3T3 fibroblasts are cultured on polystyrene dishes in DMEM and fetal bovine serum, supplemented with penicillin/streptomycmn in a 5% CO2 humidified chamber at 37 癈. Cell growth is measured by using the MTT dye reduction method.
    (Only for Reference)
Animal Research:[4]
+ Expand
  • Animal Models: Sprague Dawley female rats
  • Formulation: 4 μL of 0.9% physiological saline containing 0.02% ascorbic acid
  • Dosages: ~5 μg
  • Administration: Intrastriatal administration
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 100 mg/mL (166.93 mM)
Water 92 mg/mL (153.58 mM)
Ethanol Insoluble

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 599.03
Formula

C26 H29 Cl N2 O4 S . H3 O4 P

CAS No. 1188890-41-6
Storage powder
in solvent
Synonyms N/A

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Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID