| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
The primary target of Calcineurin substrate is calcineurin (protein phosphatase 2B, PP2B), a calcium/calmodulin-dependent serine/threonine protein phosphatase. Calcineurin regulates immune responses, cardiac hypertrophy, and neuronal function. The peptide substrate is phosphorylated by PKA and then dephosphorylated by calcineurin. The rate of dephosphorylation is proportional to calcineurin activity.
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|---|---|
| ln Vitro |
Calcineurin (PP2B) is a calcium-dependent serine/threonine phosphatase that has been linked to several biological reactions, such as the activation of lymphocytes, the development of neurons and muscles, neurite outgrowth, and the morphogenesis of vertebrate heart valves[1].
In vitro, Calcineurin substrate is used in calcineurin activity assays. The peptide is phosphorylated by PKA using [γ-32P]-ATP, and the phosphorylated peptide is incubated with calcineurin. The release of [32P]-phosphate is measured by scintillation counting. The assay is used to measure calcineurin activity in cell lysates or tissue homogenates. |
| ln Vivo |
In vivo activity data for Calcineurin substrate are not applicable, as it is an in vitro research reagent. It is not administered to animals. The peptide is used ex vivo to measure calcineurin activity in tissue samples. It serves as an analytical tool for studying calcineurin function in physiological and pathological conditions.
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| Enzyme Assay |
For non-cellular assays, Calcineurin substrate is phosphorylated by PKA in the presence of [γ-32P]-ATP. The phosphorylated peptide is purified and incubated with calcineurin in assay buffer containing calcium and calmodulin. The release of [32P]-phosphate is measured by scintillation counting. Calcineurin activity is calculated as the rate of dephosphorylation.
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| Cell Assay |
For cell-based assays, calcineurin activity in cell lysates is measured using the Calcineurin substrate. Cells are stimulated with calcium ionophore or other activators, lysed, and incubated with the phosphorylated peptide. Dephosphorylation is quantified as described above.
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| Animal Protocol |
For in vivo experiments, Calcineurin substrate is not administered to animals. It is used ex vivo to measure calcineurin activity in tissue samples. Tissues are homogenized in lysis buffer, and calcineurin activity is measured using the phosphorylated peptide substrate. The assay is used to assess calcineurin activity in animal models of disease.
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| References |
[1]. Crabtree GR, et al. Calcium, calcineurin, and the control of transcription. J Biol Chem. 2001 Jan 26;276(4):2313-6.
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| Molecular Formula |
C92H150N28O29
|
|---|---|
| Molecular Weight |
2112.3456
|
| Exact Mass |
2111.11
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| CAS # |
113873-67-9
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| Related CAS # |
Calcineurin substrate TFA
|
| PubChem CID |
16198110
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| Sequence |
Asp-Leu-Asp-Val-Pro-Ile-Pro-Gly-Arg-Phe-Asp-Arg-Arg-Val-Ser-Val-Ala-Ala-GluDLDVPIPGRFDRRVSVAAE
|
| SequenceShortening |
DLDVPIPGRFDRRVSVAAE
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| Appearance |
Typically exists as solid at room temperature
|
| LogP |
1.585
|
| Hydrogen Bond Donor Count |
32
|
| Hydrogen Bond Acceptor Count |
33
|
| Rotatable Bond Count |
69
|
| Heavy Atom Count |
149
|
| Complexity |
4690
|
| Defined Atom Stereocenter Count |
19
|
| SMILES |
CC[C@@H]([C@H](NC([C@@H]1CCCN1C([C@@H](NC([C@@H](NC([C@@H](NC([C@@H](N)CC(O)=O)=O)CC(C)C)=O)CC(O)=O)=O)C(C)C)=O)=O)C(N2CCC[C@H]2C(NCC(N[C@H](C(N[C@H](C(N[C@H](C(N[C@H](C(N[C@H](C(N[C@H](C(N[C@H](C(N[C@H](C(N[C@H](C(N[C@H](C(N[C@H](C(O)=O)CCC(O)=O)=O)C)=O)C)=O)C(C)C)=O)CO)=O)C(C)C)=O)CCCNC(N)=N)=O)CCCNC(N)=N)=O)CC(O)=O)=O)CC3=CC=CC=C3)=O)CCCNC(N)=N)=O)=O)=O)C
|
| InChi Key |
CNNRZOUCQVLOST-RBHZDPOXSA-N
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| InChi Code |
InChI=1S/C92H150N28O29/c1-13-47(10)71(118-84(143)62-28-21-35-120(62)87(146)70(46(8)9)117-81(140)59(40-67(129)130)113-78(137)56(36-43(2)3)110-74(133)51(93)38-65(125)126)88(147)119-34-20-27-61(119)83(142)103-41-63(122)106-52(24-17-31-100-90(94)95)75(134)111-57(37-50-22-15-14-16-23-50)79(138)112-58(39-66(127)128)80(139)108-53(25-18-32-101-91(96)97)76(135)107-54(26-19-33-102-92(98)99)77(136)115-69(45(6)7)86(145)114-60(42-121)82(141)116-68(44(4)5)85(144)105-48(11)72(131)104-49(12)73(132)109-55(89(148)149)29-30-64(123)124/h14-16,22-23,43-49,51-62,68-71,121H,13,17-21,24-42,93H2,1-12H3,(H,103,142)(H,104,131)(H,105,144)(H,106,122)(H,107,135)(H,108,139)(H,109,132)(H,110,133)(H,111,134)(H,112,138)(H,113,137)(H,114,145)(H,115,136)(H,116,141)(H,117,140)(H,118,143)(H,123,124)(H,125,126)(H,127,128)(H,129,130)(H,148,149)(H4,94,95,100)(H4,96,97,101)(H4,98,99,102)/t47-,48-,49-,51-,52-,53-,54-,55-,56-,57-,58-,59-,60-,61-,62-,68-,69-,70-,71-/m0/s1
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| Chemical Name |
(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-1-[(2S,3S)-2-[[(2S)-1-[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-3-carboxypropanoyl]amino]-4-methylpentanoyl]amino]-3-carboxypropanoyl]amino]-3-methylbutanoyl]pyrrolidine-2-carbonyl]amino]-3-methylpentanoyl]pyrrolidine-2-carbonyl]amino]acetyl]amino]-5-carbamimidamidopentanoyl]amino]-3-phenylpropanoyl]amino]-3-carboxypropanoyl]amino]-5-carbamimidamidopentanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-methylbutanoyl]amino]-3-hydroxypropanoyl]amino]-3-methylbutanoyl]amino]propanoyl]amino]propanoyl]amino]pentanedioic 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 |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.4734 mL | 2.3670 mL | 4.7341 mL | |
| 5 mM | 0.0947 mL | 0.4734 mL | 0.9468 mL | |
| 10 mM | 0.0473 mL | 0.2367 mL | 0.4734 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.
Products are for research use only; We do not sell to patients
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