| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
CaM-kinase II (CaM-kinase II) [IC50 = 40 nM for CaM-kinase II activity;
Ki values: 18 nM (noncompetitive with respect to syntide-2), 320 nM (competitive with respect to autocamtide-2), 10 nM (competitive with respect to CaMK-(281-289)); IC50 for autophosphorylation of CaM-kinase II = 100 nM; Inhibition of the constitutively active 30-kDa proteolytic fragment of CaM-kinase II: lowest Ki reported = 2-8 nM] [1] |
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| ln Vitro |
CaM kinase II was discovered to be effectively inhibited by autocamtide-2-related inhibitory peptide (ATP), a unique synthetic peptide that was active both in the presence and absence of Ca2+/calmodulin. Its IC50 was reported to be 40 nM. It has 500 and 50 times more potency than KN-93 and CaMK-(281-302Ala286), respectively, under the assay conditions employed [1].
AIP (40 nM IC50) inhibits CaM-kinase II activity in a concentration-dependent manner, being 50-fold more potent than CaMK-(281-302Ala286) (IC50 = 2 μM) and 500-fold more potent than KN-93 (IC50 = 20 μM). [1] AIP at 10 μM does not significantly affect the activities of PKA, PKC, or CaM-kinase IV, whereas CaMK-(281-302Ala286) at 30 μM inhibits PKC and CaM-kinase IV. [1] AIP potently inhibits autophosphorylation of CaM-kinase II with an IC50 of 100 nM. [1] Kinetic analysis shows that AIP inhibition of CaM-kinase II is noncompetitive with respect to syntide-2 (Ki = 18 nM) and competitive with respect to autocamtide-2 (Ki = 320 nM) and CaMK-(281-289) (Ki = 10 nM). [1] AIP (1 μM) completely inhibits CaM-kinase II activity regardless of calmodulin concentration (0.01-10 μM), whereas KN-93 inhibition is calmodulin-dependent and decreases with increasing calmodulin. [1] AIP (1 μM) does not affect endogenous protein phosphorylation in CaM-kinase II-depleted rat brain extracts ("P-through" and "CaM-through" preparations) in the presence or absence of Ca2+, but markedly inhibits phosphorylation induced by exogenously added purified CaM-kinase II. [1] |
| Enzyme Assay |
CaM-kinase II activity was measured in a reaction mixture containing 40 mM Hepes-NaOH (pH 8.0), 5 mM magnesium acetate, 50 μM [γ-32P]ATP, 0.01% Tween 20, 100 μM syntide-2 as substrate, 0.3 mM CaCl2, 1 μM calmodulin, and purified CaM-kinase II from rat cerebral cortex. The reaction was carried out at 30°C, and activity was determined by the phosphocellulose paper method. [1]
The activity of the active 30-kDa proteolytic fragment of CaM-kinase II was measured similarly in the presence or absence of Ca2+/calmodulin as described previously. [1] CaM-kinase IV activity was determined in the presence of 0.2 mM CaCl2 with 100 μM syntide-2 as substrate, using 40 mM Hepes-NaOH (pH 8.0) instead of 50 mM Hepes-NaOH (pH 7.0), and other conditions as previously described. [1] PKA activity was assayed by phosphate incorporation into syntide-2 in a mixture containing 40 mM Mes-NaOH (pH 7.0), 5 mM magnesium acetate, 50 μM [γ-32P]ATP, 0.01% Tween 20, 100 μM syntide-2, and the catalytic subunit of PKA from bovine heart. The reaction was initiated by addition of the catalytic subunit and activity determined by the phosphocellulose paper method. [1] PKC activity was measured in a mixture containing 40 mM Hepes-NaOH (pH 8.0), 10 mM magnesium acetate, 50 μM [γ-32P]ATP, 0.1 mM EGTA, 0.35 mM CaCl2, 0.01% Tween 20, 2.0 μg/ml 1,2-dioleoyl-rac-glycerol, 20.2 μg/ml phosphatidylserine, 100 μM syntide-2, and purified PKC from rat cerebral cortex. The reaction was started by adding PKC and activity determined by the phosphocellulose paper method. [1] For autophosphorylation of CaM-kinase II, the kinase (1.1 μg/ml) was incubated at 30°C for 10 min in a reaction mixture containing 40 mM Hepes-NaOH (pH 8.0), 5 mM magnesium acetate, 0.2 mM CaCl2, 0.1 mM EGTA, 0.01% Tween 20, 1 μM calmodulin, and 50 μM [γ-32P]ATP with varying concentrations of inhibitor. The reaction was stopped by adding 22.7 mM EDTA, and 32P incorporation into the kinase was measured by the phosphocellulose paper method. [1] |
| References | |
| Additional Infomation |
AIP is a nonphosphorylatable analog of autocamtide-2, with Ala substituted for the phosphorylation site (Thr9). [1]
The inhibition by AIP is independent of Ca2+/calmodulin, and it binds to the substrate-binding site for the autophosphorylation site, which is distinct from the exogenous substrate (e.g., syntide-2) binding site. [1] AIP does not inhibit the 30-kDa proteolytic fragment of CaM-kinase II in a calmodulin-competitive manner, unlike KN-93. [1] AIP is suggested as a useful tool for studying the physiological roles of CaM-kinase II, being more potent and selective than previously available inhibitors such as KN-93 and CaMK-(281-302Ala286). [1] |
| Molecular Formula |
C65H114N20O21
|
|---|---|
| Molecular Weight |
1822.12084984779
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| Exact Mass |
1821.07
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| CAS # |
167114-91-2
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| Related CAS # |
Autocamtide-2-related inhibitory peptide TFA
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
8.469
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| SMILES |
[C@H](CCCNC(=N)N)(C(N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](C)C(=O)N[C@H](C(=O)O)CC(C)C)=O)NC(=O)[C@H](CCCNC(=N)N)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)NC(=O)CCCCCCCCCCCCC.C(F)(F)(F)C(=O)O
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
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| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.5488 mL | 2.7441 mL | 5.4881 mL | |
| 5 mM | 0.1098 mL | 0.5488 mL | 1.0976 mL | |
| 10 mM | 0.0549 mL | 0.2744 mL | 0.5488 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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