| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Targets |
GPR54[1]; Angiogenesis[1]; The primary target of rat Kisspeptin-10 is the KISS1 receptor (KISS1R, formerly GPR54), a G protein-coupled receptor belonging to the rhodopsin family. Upon Kp-10 binding, KISS1R primarily activates the phospholipase C signaling pathway, leading to inositol trisphosphate accumulation and intracellular calcium mobilization, while also activating ERK1/2 and p38 mitogen-activated protein kinase pathways. This receptor system plays a critical role in the neuroendocrine control of the hypothalamic-pituitary-gonadal axis. As a potent vasoconstrictor and angiogenesis inhibitor, Kp-10 also exerts anti-angiogenic effects by inhibiting Sp1-mediated VEGF expression and FAK/Rho GTPase activation.
Kisspeptin-10, rat TFA targets the kisspeptin receptor (KISS1R, also known as GPR54), a G protein-coupled receptor (GPCR) that plays a critical role in the regulation of the hypothalamic-pituitary-gonadal axis. Upon binding of the peptide to GPR54, the receptor activates Gq/11 signaling, leading to phospholipase C activation, IP3-mediated calcium release, and stimulation of gonadotropin-releasing hormone (GnRH) secretion, thus controlling puberty and fertility. |
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| ln Vitro |
In vitro studies demonstrate that rat Kisspeptin-10 activates intracellular calcium responses in KISS1R-transfected CHO cells with an EC₅₀ value of 1.54–2.6 × 10⁻⁸ M. Structure-activity relationship studies reveal that Kp-10 exhibits a helicoidal structure between Asn4 and Tyr10 residues (mixed α- and 3₁₀-helix characteristics). Alanine substitutions at position 6 (phenylalanine) or position 10 (tyrosine) result in a significant increase in EC₅₀ values (>6.46 × 10⁻⁶ M), a substantial decrease in the proportion of responsive cells, and a marked increase in the time required to reach maximal response. Ala10 substitution completely disrupts the helical structure in the C-terminal region, indicating that positions 6 and 10 are critical for Kp-10 binding and functional activity at KISS1R.
In cell-free assays, Kisspeptin-10 binds to purified rat GPR54 in membrane preparations with high affinity. The Ki for displacement of radiolabeled kisspeptin is typically in the sub-nanomolar to low nanomolar range. Binding is measured by radioligand displacement assays using [125I]-kisspeptin or [3H]-kisspeptin, with bound radioactivity measured by scintillation counting after filtration. The peptide's affinity for rodent GPR54 is generally comparable to that for human GPR54. |
| ln Vivo |
Rat Kisspeptin-10 potently stimulates the hypothalamic-pituitary-gonadal axis in vivo. In rodents, Kp-10 administration significantly increases luteinizing hormone secretion, an effect mediated through stimulation of gonadotropin-releasing hormone release. Structure-activity studies show that Ala6 substitution diminishes, while Ala10 substitution completely eliminates LH secretory responses, and co-administration of these antagonist analogs does not affect the LH-releasing ability of Kp-10 itself. In a rat model of methotrexate-induced reproductive toxicity, Kisspeptin-10 (50 nmol/kg, intraperitoneal injection for 10 days) significantly ameliorates decreased sperm motility and abnormal sperm morphology, reduces malondialdehyde levels in testicular tissue, and restores reproductive organ weights. These results indicate that Kp-10 acts as a potential antioxidant to alleviate drug-induced reproductive toxicity.
In cell-based assays using HEK293 cells expressing GPR54, Kisspeptin-10, rat TFA (1-100 nM) activates calcium flux (measured by calcium-sensitive dyes such as Fluo-4 or Fura-2) and increases intracellular IP3 accumulation. The peptide also induces phosphorylation of ERK1/2 and stimulates GnRH secretion from GT1-7 GnRH neurons. The EC50 for calcium mobilization is typically in the low nanomolar range. The peptide also possesses antioxidant properties, reducing methotrexate-induced reproductive toxicity. |
| Enzyme Assay |
Cell-free receptor binding studies for rat Kisspeptin-10 primarily utilize radioligand binding assays. A typical protocol includes: 1) Prepare membrane suspensions expressing KISS1R (using native tissues or KISS1R-transfected cells); 2) Dissolve rat Kp-10 in binding buffer (containing 50 mM HEPES pH 7.4, 5 mM MgCl₂, 0.1% BSA) to prepare serial concentrations (e.g., 10⁻¹¹ to 10⁻⁶ M); 3) Add radiolabeled Kp-10 analog (e.g., [¹²⁵I]-Kp-10) and incubate for 60 minutes at room temperature; 4) Terminate the reaction by rapid vacuum filtration and wash filters with ice-cold buffer to remove unbound ligand; 5) Measure membrane-bound radioactivity using a gamma counter; 6) Calculate IC₅₀ and Kᵢ values from competition binding curves. Structure-activity relationship studies can employ alanine scanning mutagenesis to evaluate the impact of key amino acid residues on receptor binding.
The binding affinity of Kisspeptin-10 to GPR54 is measured in a cell-free radioligand binding assay. Membranes from rat or mouse cells expressing GPR54 are incubated with 0.1 nM [125I]-kisspeptin-10 and increasing concentrations (0.001-1000 nM) of unlabeled Kisspeptin-10, rat TFA. After 60 minutes at room temperature, bound and free tracer are separated by filtration through GF/B filters, and radioactivity is counted. The Ki is calculated by fitting competition curves using nonlinear regression. |
| Cell Assay |
The in vitro cell assay protocol for rat Kisspeptin-10 is as follows: 1) Seed target cells (such as CHO cells stably transfected with rat KISS1R) in culture plates and culture to appropriate density at 37°C with 5% CO₂; 2) Serum-starve cells prior to the experiment (typically using serum-free medium for 4-6 hours); 3) Stimulate cells with various concentrations of rat Kp-10 (typically 10⁻¹¹ to 10⁻⁶ M); 4) For calcium mobilization assays, label cells with fluorescent probes (e.g., Fluo-4 AM) and monitor fluorescence intensity changes in real-time after Kp-10 stimulation using a fluorescence microscope or fluorescence plate reader, recording the percentage of responsive cells and the time to reach maximal response; 5) Calculate EC₅₀ values by nonlinear regression analysis. Under typical conditions, wild-type Kp-10 reaches maximal calcium response within 2 minutes, while less active mutants require over 6 minutes.
For calcium mobilization assays, CHO or HEK293 cells stably expressing rat GPR54 are seeded in 96-well black-wall clear-bottom plates. Cells are loaded with a calcium-sensitive dye (e.g., Fluo-4 AM) for 30-60 minutes at 37degC. Varying concentrations (0.001-1000 nM) of Kisspeptin-10, rat TFA are injected automatically, and fluorescence (Ex 485/Em 525 nm) is measured in real-time using a fluorescence plate reader. The EC50 is calculated from the peak fluorescence change relative to baseline. |
| Animal Protocol |
The in vivo animal assay protocol for rat Kisspeptin-10 is as follows: 1) Use adult male rats (such as Wistar or SD rats); 2) Group assignment: randomly divide animals into vehicle control and Kp-10 treatment groups (multiple doses, e.g., 50 nmol/kg); 3) Dosing regimen: intraperitoneal injection once daily for 10 consecutive days; intravenous injection can also be used for acute pharmacodynamic studies; 4) For methotrexate-induced reproductive toxicity models, initiate Kp-10 treatment 3 days after MTX administration (single dose of 20 mg/kg intraperitoneally); 5) Sample collection: euthanize animals after treatment and collect blood, testes, epididymides, and seminal vesicles; 6) Endpoints: assess sperm motility and abnormal sperm morphology; measure malondialdehyde levels in tissue homogenates as an oxidative stress marker; weigh reproductive organs; 7) Assess hypothalamic-pituitary-gonadal axis activation by measuring serum LH and FSH levels via radioimmunoassay.
In vivo studies are performed in rats or mice. For reproductive studies, the peptide is administered intraperitoneally (e.g., 1-10 nmol/kg) to evaluate its effects on luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion. Blood samples are collected at multiple time points (0-120 minutes) post-injection, and hormone levels are measured by ELISA. For angiogenesis studies, the peptide may be administered in corneal micropocket assays or Matrigel plug assays in mice to evaluate its anti-angiogenic effects. |
| ADME/Pharmacokinetics |
Rat Kisspeptin-10 exhibits extremely rapid pharmacokinetics in vivo. Stability studies demonstrate that the decomposition half-life of this peptide in rat plasma is only 1.7 minutes at 37°C, 2.9 minutes at 25°C, and 6.8 minutes at 4°C; the principal decomposition product is the N-terminal tyrosine-deleted des-Tyr1-Kp-10. Following intravenous bolus administration of 1.0 mg/kg Kp-10, low ng/mL levels are detectable in rat plasma only during the first few minutes, becoming undetectable below the LLOQ (0.5 ng/mL) by 30 minutes post-dose. Due to the rapid clearance of the peptide in vivo, increased LH levels are commonly used as a surrogate marker for Kp-10 exposure. Rat Kp-10 has a solubility of 50 mg/mL (37.92 mM) in DMSO, and it is recommended to store the powder at -20°C for 3 years or solutions at -80°C for 6 months.
Kisspeptin-10, rat TFA has a molecular weight of 1432.46 Da and molecular formula C65H84N17O17 (plus TFA). The TFA salt improves peptide solubility. The peptide has a short plasma half-life (minutes) due to rapid proteolytic degradation, requiring frequent dosing (e.g., hourly) or continuous infusion for sustained effects. It is soluble in sterile water or PBS and should be stored as a powder at -20degC, protected from light. For in vivo use, the peptide is typically reconstituted in 0.9% saline immediately before injection. |
| Toxicity/Toxicokinetics |
According to the Material Safety Data Sheet, rat Kisspeptin-10 is classified as a non-hazardous substance or mixture with no GHS hazard classification identified. This product is for research use only and is not intended for human or veterinary use. In animal studies, no significant toxicity has been observed following intraperitoneal administration of 50 nmol/kg for 10 consecutive days, with animals showing good tolerance. In the methotrexate-induced reproductive toxicity model, Kp-10 treatment not only showed no toxicity but also alleviated MTX-induced reproductive damage as a potential antioxidant. For handling and storage, avoid inhalation and contact with skin, eyes, and clothing; appropriate personal protective equipment (such as rubber gloves and protective clothing) is recommended, and operations should be performed in well-ventilated areas.
Kisspeptin-10 is well-tolerated at typical research doses (e.g., 1-10 nmol/kg) in rodents. No significant systemic toxicity has been reported, although careful monitoring of blood pressure is advised due to its vasoconstrictor properties. At higher doses, transient changes in heart rate and blood pressure may occur. Reproductive effects include activation of the hypothalamic-pituitary-gonadal axis. The peptide is for research use only; no clinical safety data are available. |
| References |
[1]. Ameliorating effect of kisspeptin-10 on methotrexate-induced sperm damages and testicular oxidative stress in rats. Andrologia. 2018 Oct;50(8):e13057.
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| Additional Infomation |
Kisspeptin-10, rat TFA is a research-use-only peptide that serves as a potent agonist for the rat/mouse kisspeptin receptor (GPR54). It is used in studies of puberty onset, fertility regulation, reproductive endocrinology, neuroendocrinology, and cancer biology (as a metastasis suppressor). The peptide has potential applications in treating reproductive disorders and protecting against chemotherapy-induced gonadal toxicity. It is not approved for clinical use.
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| Molecular Formula |
C65H84F3N17O17
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|---|---|
| Molecular Weight |
1432.46
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| Exact Mass |
1431.6183187
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| Related CAS # |
Kisspeptin-10, rat;478507-53-8
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| Appearance |
Typically exists as solid at room temperature
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| Hydrogen Bond Donor Count |
20
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| Hydrogen Bond Acceptor Count |
22
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| Rotatable Bond Count |
38
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| Heavy Atom Count |
102
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| Complexity |
2710
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| Defined Atom Stereocenter Count |
9
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| SMILES |
CC(C)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N)NC(=O)CNC(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC3=CNC4=CC=CC=C43)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC5=CC=C(C=C5)O)N.C(=O)(C(F)(F)F)O
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| InChi Key |
ZKCRYTJFHFFTIK-QDUZOICUSA-N
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| InChi Code |
InChI=1S/C63H83N17O15.C2HF3O2/c1-33(2)23-45(58(91)74-43(13-8-22-70-63(68)69)57(90)75-44(54(67)87)25-36-16-20-39(83)21-17-36)73-53(86)31-72-56(89)46(26-34-9-4-3-5-10-34)77-62(95)50(32-81)80-61(94)49(29-52(66)85)79-59(92)47(27-37-30-71-42-12-7-6-11-40(37)42)78-60(93)48(28-51(65)84)76-55(88)41(64)24-35-14-18-38(82)19-15-35;3-2(4,5)1(6)7/h3-7,9-12,14-21,30,33,41,43-50,71,81-83H,8,13,22-29,31-32,64H2,1-2H3,(H2,65,84)(H2,66,85)(H2,67,87)(H,72,89)(H,73,86)(H,74,91)(H,75,90)(H,76,88)(H,77,95)(H,78,93)(H,79,92)(H,80,94)(H4,68,69,70);(H,6,7)/t41-,43-,44-,45-,46-,47-,48-,49-,50-;/m0./s1
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| Chemical Name |
(2S)-N-[(2S)-1-[[(2S)-4-amino-1-[[(2S)-1-[[(2S)-1-[[2-[[(2S)-1-[[(2S)-1-[[(2S)-1-amino-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-2-oxoethyl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-1,4-dioxobutan-2-yl]amino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]-2-[[(2S)-2-amino-3-(4-hydroxyphenyl)propanoyl]amino]butanediamide;2,2,2-trifluoroacetic acid
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| Synonyms |
Kisspeptin-10, rat (TFA); KISSPEPTIN-10, RAT TFA; HY-P1197A; Kisspeptin-10 (mouse, rat) (TFA); MS-32166; .
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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) |
DMSO :≥ 50 mg/mL (~34.90 mM)
H2O :~10 mg/mL (~6.98 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (1.75 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (1.75 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (1.75 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.6981 mL | 3.4905 mL | 6.9810 mL | |
| 5 mM | 0.1396 mL | 0.6981 mL | 1.3962 mL | |
| 10 mM | 0.0698 mL | 0.3490 mL | 0.6981 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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