| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Targets |
The primary target of Autocamtide 2 is calcium/calmodulin-dependent protein kinase II (CaMKII). CaMKII is a serine/threonine protein kinase that is activated by the binding of calcium-bound calmodulin (Ca²⁺/CaM). Upon activation, CaMKII undergoes autophosphorylation at Thr286 (for the α isoform), which renders the kinase partially calcium-independent and sustains its activity even after calcium levels decline. CaMKII plays a pivotal role in the central nervous system, where it is essential for synaptic plasticity, learning, and memory. It phosphorylates a diverse array of substrates, including ion channels, neurotransmitter receptors, transcription factors, and other kinases, thereby regulating neuronal excitability, synaptic strength, and gene expression. In addition to its role in the brain, CaMKII is also involved in the regulation of cardiac function, smooth muscle contraction, and immune responses. Dysregulation of CaMKII activity has been implicated in various diseases, including cardiac hypertrophy, heart failure, and cancer. Autocamtide 2 is a specific substrate for CaMKII and is not phosphorylated by other kinases, making it an ideal tool for measuring CaMKII activity in complex biological samples.
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| ln Vitro |
Ca/calmodulin-dependent protein kinase (CaMK) is a multifunctional serine/threonine kinase whose activity is regulated by Ca2+ signaling. It is flexible and known for its vital function in learning and memory. Recent studies have demonstrated that high amounts of several CaMK isoforms, especially CaMKII, are expressed in a variety of malignancies, such as lung, breast, prostate, and colon cancers. CaMKII phosphorylates roughly 40 different proteins, including enzymes, ion channels, kinases, and transcription factors, and plays a vital role in controlling the proliferation, differentiation, and survival of a variety of cancer cells [1].
Autocamtide 2 is a highly selective peptide substrate for CaMKII, and its primary in vitro activity is its phosphorylation by the kinase. The peptide sequence is derived from the autophosphorylation site of CaMKII and is specifically recognized and phosphorylated by the kinase. In kinase activity assays, Autocamtide 2 is incubated with a sample containing CaMKII (e.g., purified enzyme, cell lysates, or tissue extracts) in the presence of ATP and Ca²⁺/calmodulin. The incorporation of phosphate into the peptide is then measured using various methods, such as radioactive counting (using γ-³²P-ATP), fluorescence polarization, or mass spectrometry. The amount of phosphorylated peptide is directly proportional to the activity of CaMKII in the sample. The selectivity of Autocamtide 2 ensures that the measured activity is specific to CaMKII, minimizing interference from other kinases. The compound is supplied as a white to off-white solid powder with a purity of ≥98%. |
| ln Vivo |
In vivo, Autocamtide 2 is not used as a therapeutic agent but rather as a research tool. Its primary application is in the measurement of CaMKII activity in tissue extracts or cell lysates obtained from animal models. By measuring CaMKII activity using Autocamtide 2, researchers can assess the activation state of the kinase in various physiological and pathological conditions, such as during learning and memory formation, in response to stress or injury, or in disease models. Changes in CaMKII activity can be correlated with behavioral, biochemical, or histological endpoints to gain insights into the role of CaMKII in these processes. The compound's high selectivity makes it a valuable tool for such studies.
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| Enzyme Assay |
In vitro enzyme assays for Autocamtide 2 are designed to measure the activity of CaMKII. In a typical assay, Autocamtide 2 is incubated with a CaMKII-containing sample in a reaction buffer containing ATP (often radiolabeled with ³²P), magnesium chloride, and calcium/calmodulin. The reaction is allowed to proceed for a set period, and then the reaction is stopped, and the phosphorylated peptide is separated from the unreacted ATP. The amount of ³²P incorporated into the peptide is quantified using scintillation counting. Alternatively, non-radioactive methods, such as fluorescence-based or luminescence-based assays, can be used. The kinase activity is calculated from the amount of phosphorylated peptide produced per unit of time. Inhibitors or activators of CaMKII can be included in the assay to study their effects on the kinase.
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| Cell Assay |
In vitro cell-based assays using Autocamtide 2 are less common, as the substrate is typically used in cell-free assays. However, cellular activity of CaMKII can be assessed by treating cells with stimuli that activate the kinase (e.g., ionomycin or glutamate) and then preparing cell lysates. The lysates are then used in the in vitro kinase assay with Autocamtide 2 to measure CaMKII activity. Alternatively, the phosphorylation of endogenous CaMKII substrates can be measured by western blotting using phospho-specific antibodies. These approaches allow researchers to link cellular stimulation to CaMKII activation.
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| Animal Protocol |
In vivo animal studies using Autocamtide 2 are not typically performed, as the compound is not administered to animals. Instead, tissues or cells from animals are collected, and CaMKII activity is measured ex vivo using Autocamtide 2 in in vitro kinase assays. This approach is used to study changes in CaMKII activity in various disease models, such as models of Alzheimer's disease, cardiac hypertrophy, or cancer. By correlating changes in CaMKII activity with disease progression or treatment response, researchers can gain insights into the role of CaMKII in these conditions.
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| ADME/Pharmacokinetics |
Autocamtide 2 is a peptide with a molecular weight of approximately 1527.77 and a molecular formula of C₆₅H₁₁₈N₂₂O₂₀. It is supplied as a white to off-white solid powder with a purity of ≥98%. The compound is soluble in aqueous buffers and is typically stored as a powder at -20°C for long-term stability. For use in kinase assays, it is dissolved in an appropriate buffer to prepare a stock solution. The compound's high selectivity for CaMKII makes it a valuable tool for studying the role of this kinase in various biological processes and diseases.
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| Toxicity/Toxicokinetics |
The toxicological profile of Autocamtide 2 is not extensively documented, as it is a peptide substrate used in research rather than a therapeutic agent. At the concentrations used in kinase assays, it is not expected to be toxic. For laboratory handling, standard safety precautions for research chemicals should be observed, including the use of personal protective equipment (gloves, lab coat, safety goggles) and working in a well-ventilated area. The compound is intended for research use only and is not for human therapeutic or diagnostic use.
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| References | |
| Additional Infomation |
Autocamtide 2 is a highly selective peptide substrate for calcium/calmodulin-dependent protein kinase II (CaMKII). It has a molecular formula of C₆₅H₁₁₈N₂₂O₂₀ and a molecular weight of approximately 1527.77. The compound is supplied as a white to off-white solid powder with a purity of ≥98%. It is soluble in aqueous buffers and is typically stored at -20°C. Autocamtide 2 is a valuable tool for studying CaMKII activity and function in various biological systems, including neuroscience, cardiovascular research, and cancer biology. Its high selectivity ensures that the measured kinase activity is specific to CaMKII, making it an essential reagent for researchers studying this important signaling pathway.
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| Molecular Formula |
C65H118N22O20
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|---|---|
| Molecular Weight |
1527.76782
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| Exact Mass |
1526.89
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| CAS # |
129198-88-5
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| PubChem CID |
137700269
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| Appearance |
White to off-white solid powder
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| LogP |
2.648
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| Hydrogen Bond Donor Count |
25
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| Hydrogen Bond Acceptor Count |
30
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| Rotatable Bond Count |
55
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| Heavy Atom Count |
114
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| Complexity |
3120
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| Defined Atom Stereocenter Count |
14
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| SMILES |
C[C@H]([C@@H](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](CC(C)C)C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)N)O.C(=O)(C(F)(F)F)O
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| InChi Key |
LLNOFJJCOGGKCB-ZOUHFXOGSA-N
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| InChi Code |
InChI=1S/C65H118N22O20.C2HF3O2/c1-31(2)28-43(83-51(94)34(7)76-54(97)38(17-11-13-25-67)78-53(96)37(68)16-10-12-24-66)60(103)80-40(19-15-27-75-65(72)73)55(98)79-39(18-14-26-74-64(70)71)56(99)81-41(20-22-46(69)89)57(100)82-42(21-23-47(90)91)58(101)87-50(36(9)88)62(105)86-49(33(5)6)61(104)84-44(30-48(92)93)59(102)77-35(8)52(95)85-45(63(106)107)29-32(3)4;3-2(4,5)1(6)7/h31-45,49-50,88H,10-30,66-68H2,1-9H3,(H2,69,89)(H,76,97)(H,77,102)(H,78,96)(H,79,98)(H,80,103)(H,81,99)(H,82,100)(H,83,94)(H,84,104)(H,85,95)(H,86,105)(H,87,101)(H,90,91)(H,92,93)(H,106,107)(H4,70,71,74)(H4,72,73,75);(H,6,7)/t34-,35-,36+,37-,38-,39-,40-,41-,42-,43-,44-,45-,49-,50-;/m0./s1
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| Chemical Name |
(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S,3R)-2-[[(2S)-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-6-amino-2-[[(2S)-2,6-diaminohexanoyl]amino]hexanoyl]amino]propanoyl]amino]-4-methylpentanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]-5-oxopentanoyl]amino]-4-carboxybutanoyl]amino]-3-hydroxybutanoyl]amino]-3-methylbutanoyl]amino]-3-carboxypropanoyl]amino]propanoyl]amino]-4-methylpentanoic acid;2,2,2-trifluoroacetic acid
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 Vitro) |
H2O : ~50 mg/mL (~32.73 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (65.45 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.6545 mL | 3.2727 mL | 6.5455 mL | |
| 5 mM | 0.1309 mL | 0.6545 mL | 1.3091 mL | |
| 10 mM | 0.0655 mL | 0.3273 mL | 0.6545 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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