| Targets |
PKA RII peptide TFA targets protein kinase A (PKA) as a substrate, but it is not an inhibitor; rather, it is used to measure PKA activity or as a substrate for subsequent detection of calcineurin. For calcineurin assays, the phosphorylated peptide serves as a substrate for the phosphatase, allowing quantification of calcineurin activity.
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| ln Vitro |
PKA RII peptide TFA itself has no direct biological activity. It is a substrate peptide that, when incubated with PKA in the presence of ATP, becomes phosphorylated on the serine residue (typically Ser15 or Ser16 in the sequence). The phosphorylated peptide is then used to detect calcineurin activity by measuring the release of free phosphate or using a coupled enzyme assay. The peptide is not cytotoxic or pharmacologically active.
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| ln Vivo |
No direct in vivo activity is reported for PKA RII peptide TFA, as it is a biochemical tool for in vitro assays. The peptide is used in cell‑free systems or cell lysates to measure endogenous PKA or calcineurin activity, but it is not administered systemically as a therapeutic or research agent. Its utility lies in quantifying enzyme activity in tissue or cell extracts.
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| Enzyme Assay |
PKA RII peptide TFA is evaluated by a calcineurin phosphatase assay using the phosphorylated form of the peptide. In a typical experiment, the peptide is first phosphorylated by PKA in the presence of ATP. After purifying the phosphorylated peptide, it is incubated with purified calcineurin or a cell lysate containing calcineurin in phosphatase buffer. The release of free phosphate is measured by a malachite green or biomol green assay, or using a coupled enzyme system that produces a colorimetric or fluorescent signal.
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| Cell Assay |
PKA RII peptide TFA is not used in direct cell‑based activity assays. Instead, cell lysates containing PKA or calcineurin are used as enzyme sources. Cells (e.g., HEK293, Neuro2a) are lysed in extraction buffer, and the lysate is cleared by centrifugation. For PKA activity measurement, the lysate is incubated with the peptide, ATP, and a PKA‑specific buffer, and phosphate incorporation is measured. For calcineurin activity, the lysate is incubated with the pre‑phosphorylated peptide in the presence of calcium and calmodulin.
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| Animal Protocol |
PKA RII peptide TFA is not used directly in animal experiments. However, tissue lysates from animals (e.g., mouse brain, heart, liver) can be prepared and used in vitro for PKA or calcineurin activity assays as described above. For example, mice may be treated with pharmacological agents to modulate PKA or calcineurin activity, then tissues are harvested, lysed, and the peptide is used to measure the activity of these enzymes in the lysates.
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| ADME/Pharmacokinetics |
PKA RII peptide TFA (C₉4H1₅1F3N2₈O31, MW = 2226.37) is a 19‑amino acid peptide with the sequence DLDVPIPGRFDRRVSVAAE. It is soluble in water (H2O) at 25 mg/mL (11.23 mM). The powder should be stored sealed, away from moisture, at ‑80degC for up to 2 years or at ‑20degC for up to 1 year. Stock solutions are stable for 6 months at ‑80degC or 1 month at ‑20degC. No in vivo PK data are applicable.
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| Toxicity/Toxicokinetics |
No toxicity data are reported for PKA RII peptide TFA. As a synthetic peptide used as a biochemical tool, it is not intended for human or veterinary use. Standard laboratory safety precautions should be followed when handling peptides, including avoiding inhalation, ingestion, and skin contact. Peptides are generally considered to have low toxicity at the concentrations used in enzyme assays (micromolar range).
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| References | |
| Additional Infomation |
PKA RII peptide TFA is a well‑established biochemical tool for studying PKA and calcineurin signaling pathways. It is derived from the regulatory subunit II of protein kinase A and contains the canonical PKA phosphorylation site (Arg‑Arg‑X‑Ser). After phosphorylation, it serves as a specific substrate for calcineurin (protein phosphatase 2B). The peptide is for research use only and has no clinical or therapeutic applications. It has not received regulatory approval for any indication.
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| Appearance |
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 : 25 mg/mL (11.23 mM; with sonication)
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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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