| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Autocamtide‑3 targets the active site of CaMKII, specifically recognising the CaMKII substrate consensus sequence (RXXS/T). It is phosphorylated by CaMKII on a specific serine residue. The peptide has high selectivity for CaMKII over other kinases, including PKA, PKC, CaMKIV and CaMKI, making it an excellent tool for measuring CaMKII activity in complex biological samples.
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| ln Vitro |
In a 50 μL kinase test procedure, 20 μM Autocamtide-3 acetate (H2N-Lys-Lys-Ala-Leu-His-Arg-Gln-Glu-Thr-Val-Asp-Ala-Leu-COOH) and 5 μL of CaMKII are utilized. Catalytic activity is shown by autocamtide-3 acetate[1].
In cell‑free kinase assays, autrocamtide‑3 acetate is phosphorylated by recombinant CaMKII in a Ca2+/calmodulin‑dependent manner. The Km value for CaMKII is typically 5‑15 uM, and the kcat is in the range of 1‑5 s-¹, making it a superior substrate compared to generic peptides like syntide‑2. The peptide is not significantly phosphorylated by PKA, PKC, CaMKIV or other basophilic kinases at concentrations up to 100 uM. |
| ln Vivo |
No direct in vivo activity is described, as autocamtide‑3 is a substrate rather than a therapeutic agent. However, it is used in ex vivo kinase activity assays to measure CaMKII activation in tissue lysates from animal models of neurological disorders, cardiac hypertrophy and synaptic plasticity. The peptide helps to elucidate the role of CaMKII in learning and memory, ischaemic injury and heart failure.
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| Enzyme Assay |
A standard CaMKII kinase assay (radioactive): Autocamtide‑3 acetate (final concentration 10‑200 uM) is incubated with recombinant CaMKII (5‑20 nM) in assay buffer (50 mM HEPES pH 7.4, 10 mM MgCl2, 0.5 mM CaCl2, 1 uM calmodulin, 1 mM DTT, 0.1 mg/mL BSA) for 10 min at 30degC. The reaction is initiated by adding 100 uM ATP (with 0.5 uCi/well 33P‑ATP). After 5‑20 min, the reaction is spotted onto P81 phosphocellulose paper, washed 3 times with 0.75% phosphoric acid, and counted by liquid scintillation. Controls without Ca2+/calmodulin or with EGTA (2 mM) determine Ca2+‑independent activity. IC50 values for CaMKII inhibitors can be calculated by adding inhibitors to the reaction mix.
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| Cell Assay |
A general cellular CaMKII activity assay (ex vivo): Cells or tissue samples (e.g., primary neurons, cardiac myocytes, brain tissue lysates) are lysed in RIPA buffer containing phosphatase and protease inhibitors. Protein concentration is normalised (0.5‑1 ug/uL). Autocamtide‑3 acetate (50 uM) is added to the lysate along with Ca2+/calmodulin (0.5 mM CaCl2, 1 uM calmodulin) and ATP (100 uM) in assay buffer. The reaction is incubated at 30degC for 30 min. Phosphorylation is quantified by transferring the reaction to a streptavidin‑coated plate (if biotinylated peptide is used) and detecting with an anti‑phospho‑specific antibody or by measuring incorporated 33P‑ATP. CaMKII activity is expressed as pmol phosphate incorporated per mg protein per minute.
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| Animal Protocol |
A general animal protocol for CaMKII activity measurement: Male C57BL/6 mice or Sprague‑Dawley rats are subjected to treatments that modulate CaMKII activity (e.g., induction of long‑term potentiation (LTP) by electrical stimulation, ischaemia‑reperfusion injury, cardiac pressure overload by transverse aortic constriction). Animals are euthanised by cervical dislocation or isoflurane overdose. Brain (hippocampus, cortex) or cardiac tissue is rapidly dissected, snap‑frozen in liquid nitrogen, and stored at -80degC. For activity measurement, tissue is homogenised in lysis buffer and processed as described in the cellular assay protocol. Enzyme activity is normalised to total protein content (BCA assay) or to total CaMKII protein level (western blot).
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| ADME/Pharmacokinetics |
As a peptide substrate, autocamtide‑3 acetate has a short plasma half‑life (minutes) due to rapid proteolytic degradation and is not used as a therapeutic agent. For ex vivo assays, the peptide is added directly to tissue lysates, so pharmacokinetic properties are not applicable. The acetate salt provides good water solubility (>10 mg/mL in water and physiological buffers). For storage, the lyophilised powder should be kept at -20degC or -80degC, protected from moisture and light.
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| Toxicity/Toxicokinetics |
Autocamtide‑3 acetate is a research‑grade peptide and is not intended for human or therapeutic use. At the concentrations used in kinase assays (10‑200 uM), the peptide is non‑toxic to cells. For animal studies, the compound is not directly administered; only the tissue lysates are assayed. Standard laboratory safety practices (gloves, lab coat, eye protection) should be used when handling the lyophilised powder or stock solutions.
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| References | |
| Additional Infomation |
Autocamtide‑3 has the peptide sequence KKALRRQEAVDAL (with the phosphorylation site typically at the serine within the RXXS/T motif; the exact sequence may vary by manufacturer). It is an improved version of autocamtide‑2, with higher selectivity and catalytic efficiency for CaMKII. The peptide is acetylated at the N‑terminus and amidated at the C‑terminus to prevent proteolysis and improve stability. Autocamtide‑3 is widely used in neuroscience and cardiovascular research to study CaMKII‑dependent signalling pathways, including synaptic plasticity, neuronal excitability, cardiac contractility and hypertrophy.
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| Molecular Formula |
C67H117N21O22.XC2H4O2
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|---|---|
| Molecular Weight |
1508.72 (free acid)
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| Related CAS # |
Autocamtide-3;142828-10-2
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| Appearance |
White to off-white solid powder
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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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 :~100 mg/mL
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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.) |
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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