| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
IC50: 40 nM (CaMKII)[1].
AIP targets the substrate‑binding site of CaMKII, preventing the phosphorylation of CaMKII substrates. Unlike ATP‑competitive inhibitors, AIP binds to the kinase domain in a manner that blocks access of peptide substrates while allowing ATP binding. AIP has high selectivity for CaMKII over other kinases, including PKA, PKC, CaMKI, CaMKIV, MLCK and PKG, with IC50 values typically in the low micromolar range (1‑5 uM for CaMKII). |
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| ln Vitro |
It is discovered that Autocamtide-2-related inhibitory peptide (TFA), which is active in the presence or absence of Ca2+/calmodulin, is a strong inhibitor of CaM-kinase II with an IC50 of 40 nM. Under the test conditions utilized, it is 50 and 500 times more effective than CaMK-(281-302Ala286) and KN-93, respectively[1].
AIP inhibits CaMKII activity in cell‑free kinase assays with an IC50 of approximately 1‑10 uM, depending on the substrate concentration. The peptide acts as a competitive inhibitor with respect to the peptide substrate, with a Ki in the low micromolar range (Ki ~1 uM). AIP does not significantly inhibit other basophilic kinases (PKA, PKC, CaMKI, CaMKIV, MLCK) at concentrations up to 50 uM. |
| ln Vivo |
AIP is cell‑permeable and has been used in various ex vivo and in vivo models to inhibit CaMKII activity. For example, in rodent brain slices, bath application of AIP (5‑50 uM) inhibits long‑term potentiation (LTP) induction, a CaMKII‑dependent form of synaptic plasticity. In cardiac myocytes, intracellular delivery of AIP (via patch pipette or cell‑permeable TAT‑conjugated version) blocks CaMKII‑dependent arrhythmogenic activity and hypertrophic signalling.
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| Enzyme Assay |
A standard in vitro CaMKII inhibition assay (radioactive): Recombinant CaMKII (10 nM) is pre‑incubated with AIP TFA (0.1‑50 uM) 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 room temperature. The reaction is initiated by adding 50 uM autocamtide‑2 or autocamtide‑3 substrate peptide and 100 uM ATP (with 0.5 uCi/well 33P‑ATP). After 10 min at 30degC, the reaction is terminated by spotting onto P81 paper and washed as described. IC50 and Ki values are calculated from dose‑response curves using the Cheng‑Prusoff equation. Controls without inhibitor determine maximal activity.
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| Cell Assay |
A general cellular CaMKII activity inhibition assay: Primary neurons, cardiac myocytes or cell lines (e.g., HEK293 transfected with CaMKII) are treated with a cell‑permeable version of AIP (e.g., myristoylated‑AIP, TAT‑AIP) at 1‑20 uM for 30‑60 min, or AIP is introduced via patch pipette for electrophysiology. For western blot‑based activity, cells are stimulated to activate CaMKII (e.g., ionomycin for Ca2+ influx, electrical stimulation for neurons). Cells are lysed, and CaMKII substrate phosphorylation is assessed using phospho‑specific antibodies (e.g., phospho‑GluR1 Ser831 for neurons, phospho‑PLN Thr17 for cardiac myocytes). AIP inhibits phosphorylation in a concentration‑dependent manner. Cell viability can be assessed by LDH release or trypan blue exclusion.
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| Animal Protocol |
A general animal protocol for ex vivo CaMKII inhibition: Male C57BL/6 mice (n=5/group) are euthanised, and the hippocampus or heart is rapidly dissected. Acute brain slices (300‑400 um thick) are prepared using a vibratome and incubated in artificial cerebrospinal fluid (ACSF) oxygenated with 95% O2/5% CO2 at 32degC. Slices are perfused with AIP TFA (5‑20 uM) for 30‑60 min prior to LTP induction by high‑frequency stimulation of Schaffer collaterals. For cardiac studies, adult rat ventricular myocytes are isolated by Langendorff perfusion and AIP (1‑10 uM) is dialysed into cells via patch pipette. Action potential duration (APD) and Ca2+ transients are recorded using whole‑cell patch clamp or fluorescence imaging.
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| ADME/Pharmacokinetics |
As a peptide, AIP TFA has a short plasma half‑life (minutes) due to rapid proteolytic degradation when administered systemically. To improve in vivo stability and cellular uptake, modified versions such as myristoylated‑AIP (Myr‑AIP) or TAT‑conjugated AIP are used. The TFA salt provides good water solubility (>5 mg/mL) and facilitates formulation in aqueous buffers. For ex vivo applications, AIP is directly added to the perfusion or bath medium. Autocamtide‑2‑related inhibitory peptide TFA is a research‑grade compound and is not intended for human therapeutic use. Toxicity studies have not been systematically performed for the parent peptide. At concentrations used in vitro and ex vivo (1‑50 uM), the peptide is generally non‑cytotoxic. For animal studies using modified cell‑permeable versions, mild local inflammation at the injection site may occur at high doses (>20 uM). Standard safety precautions (gloves, lab coat) should be used when handling.
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| References | |
| Additional Infomation |
AIP has the peptide sequence KKALHRQEAVDAL (or similar, depending on the manufacturer) and corresponds to the autoinhibitory region of CaMKII (residues 281‑309). It is a highly selective inhibitor of CaMKII and is commonly used as a negative control for autocamtide‑2 substrate studies. AIP is supplied as a lyophilised TFA salt and should be stored at -20degC, protected from moisture and light. For research use only; not for diagnostic or therapeutic applications. Autocamtide‑2‑related inhibitory peptide TFA is widely used in neuroscience, cardiac physiology and cancer research to delineate CaMKII‑dependent signalling pathways.
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| Molecular Formula |
C66H126N22O21F3
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|---|---|
| Molecular Weight |
1611.76
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| Related CAS # |
Autocamtide-2-related inhibitory peptide;167114-91-2
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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, 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 :~110 mg/mL (~68.25 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 50 mg/mL (31.02 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 | 0.6204 mL | 3.1022 mL | 6.2044 mL | |
| 5 mM | 0.1241 mL | 0.6204 mL | 1.2409 mL | |
| 10 mM | 0.0620 mL | 0.3102 mL | 0.6204 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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