2. GPCR/G Protein
  3. Neuropeptide FF Receptor
  4. Neuropeptide FF

Neuropeptide FF (NPFF), an octapeptide belonging to the RF-amide family of peptides, is a NPFF1 and NPFF2 receptors agonist with Ki values of 2.82 nM and 0.21 nM, respectively. Neuropeptide FF induces abstinence syndrome, exerts antiopioid and analgesic effects, releases via calcium-dependent mechanisms from rat spinal cord, regulates memory, autonomic function, and neuroendocrine function, modulates pain and opioid antinociception, reduces food intake, stimulates water intake, alters cardiovascular parameters, and shows differential activity in hypothalamic paraventricular nucleus neurons. Neuropeptide FF is present in mammalian central nervous system and periphery, with NPFF-immunoreactivity increases in rat cerebrospinal fluid during opiate tolerance, and its NPFF gene and NPFF-R2 gene are up-regulated in rat spinal cord and dorsal root ganglia during peripheral inflammation. Neuropeptide FF can be used for the research of opioid tolerance, morphine-induced analgesia, abstinence syndrome, pain, hypertension, nociception, inflammatory pain, and neuropathic pain.

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Neuropeptide FF

Neuropeptide FF Chemical Structure

CAS No. : 99566-27-5

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Based on 1 publication(s) in Google Scholar

Neuropeptide FF Related Antibodies

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Description

Neuropeptide FF (NPFF), an octapeptide belonging to the RF-amide family of peptides, is a NPFF1 and NPFF2 receptors agonist with Ki values of 2.82 nM and 0.21 nM, respectively. Neuropeptide FF induces abstinence syndrome, exerts antiopioid and analgesic effects, releases via calcium-dependent mechanisms from rat spinal cord, regulates memory, autonomic function, and neuroendocrine function, modulates pain and opioid antinociception, reduces food intake, stimulates water intake, alters cardiovascular parameters, and shows differential activity in hypothalamic paraventricular nucleus neurons. Neuropeptide FF is present in mammalian central nervous system and periphery, with NPFF-immunoreactivity increases in rat cerebrospinal fluid during opiate tolerance, and its NPFF gene and NPFF-R2 gene are up-regulated in rat spinal cord and dorsal root ganglia during peripheral inflammation. Neuropeptide FF can be used for the research of opioid tolerance, morphine-induced analgesia, abstinence syndrome, pain, hypertension, nociception, inflammatory pain, and neuropathic pain[1][2][3][4][5][6][7].

IC50 & Target[7]

NPFF1-R

2.82 nM (Ki)

NPFF2-R

0.21 nM (Ki)

In Vitro

Neuropeptide FF (0.1 pM-100 nM) exerts concentration-dependent effects on human peripheral blood lymphocyte proliferation, enhancing proliferation at 0.1 pM and inhibiting proliferation at 100 nM[1].
Neuropeptide FF binds to two distinct sites on human Jurkat T lymphocyte cells, with a high-affinity site (Kd of 0.29 nM) and a low-affinity site (Kd of 0.512 μM)[1].
Neuropeptide FF binds with high affinity to both human NPFF-R1 and NPFF-R2 expressed in CHO-NFA-bla cells, with a slightly higher affinity for NPFF-R2 (IC50 = 3.1 nM) compared to NPFF-R1 (IC50 = 6.2 nM)[3].
Neuropeptide FF specifically potentiates acid-gated currents in Xenopus oocytes expressing heteromeric human ASIC2a + ASIC3 channels by delaying channel desensitization[3].
Neuropeptide FF reduces depolarization-induced intracellular calcium elevation in dissociated mouse dorsal root ganglion neurons[3].
Neuropeptide FF (NPFF) (0.01-10 μM; 5 min) dose-dependently activates ERK phosphorylation in dSH-SY5Y cells via a PKA-, inducible nitric oxide synthase-, and neuronal nitric oxide synthase-dependent mechanism, with peak activation at 10 μM for 5 min[4].
Neuropeptide FF (NPFF) (0.01-10 μM; 90-135 min) activates the upstream components of the NF-κB pathway in dSH-SY5Y cells in a time- and dose-dependent, ERK-independent manner, with peak IKKα/β phosphorylation at 10 μM for 120 min and peak I-κKα phosphorylation at 10 μM for 135 min[4].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Neuropeptide FF Related Antibodies

Western Blot Analysis[4]

Cell Line: Differentiated human neuroblastoma dSH-SY5Y cells
Concentration: 0.01 μM, 0.1 μM, 1 μM, 10 μM
Incubation Time: 5 min (direct treatment); 2 h (inhibitor pre-incubation) followed by 5 min (NPFF treatment)
Result: Stimulated ERK phosphorylation in a dose-dependent manner, with peak response at 10 μM.

Western Blot Analysis[4]

Cell Line: Differentiated human neuroblastoma dSH-SY5Y cells
Concentration: 10 μM
Incubation Time: 90 min, 105 min, 120 min, 135 min
Result: Induced time-dependent phosphorylation of IKKα/β, with increased phosphorylation observed at 90 min, peak activation at 120 and 135 min.
Induced dose-dependent phosphorylation of IKKα/β after 120 min of treatment.
Induced time-dependent phosphorylation of I-κKα, with increased phosphorylation at 90 min, peak activation at 135 min.
In Vivo

Neuropeptide FF (intracerebroventricular; intrathecal; focal brainstem microinjection) modulates cardiovascular function via dose-dependent changes in blood pressure, heart rate, and selective activation/inhibition of hypothalamic and brainstem neurons, and blunts hypovolemia-induced vasopressin release in rats[2].
Neuropeptide FF (intrathecal) produces potent antiallodynia in rats with carrageenan-induced inflammatory nociception[3].
Neuropeptide FF (intrathecal) produces potent antiallodynia in rats with mononeuropathic or diabetic neuropathic nociception[3].
Neuropeptide FF (NPFF) (5-10 μg/rat; i.c.v.; single injection) produces a dose-related reduction in food intake during the first 60 minutes in 24-hour food-deprived male Sprague-Dawley rats, with the 10 μg/rat dose inducing a statistically significant decrease[5].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model: Sprague-Dawley (male, 250-270 g, 24-hour food deprivation)[5]
Dosage: 5 μg/rat; 10 μg/rat
Administration: i.c.v.; single injection
Result: Reduced food intake in the first 60 minutes after injection in a dose-related manner.
Produced a statistically significant reduction in food intake relative to control at 10 μg/rat.
Showed no significant differences in cumulative food intake at 2 hours after injection or later.
Molecular Weight

1081.27

Formula

C54H76N14O10
CAS No.
Appearance

Solid

Color

White to off-white

Sequence

Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2
Sequence Shortening

FLFQPQRF-NH2
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

Sealed storage, away from moisture and light

Powder -80°C 2 years
-20°C 1 year

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

Solvent & Solubility
In Vitro: 

DMSO : 100 mg/mL (92.48 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H2O : 50 mg/mL (46.24 mM; Need ultrasonic)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 0.9248 mL 4.6242 mL 9.2484 mL
5 mM 0.1850 mL 0.9248 mL 1.8497 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

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Concentration
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Volume
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Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

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Volume (start)

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Concentration (final)

C2

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V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.5 mg/mL (2.31 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.5 mg/mL (2.31 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Calculation results:
Working solution concentration: mg/mL
This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only.If necessary, please contact MedChemExpress (MCE).
Purity & Documentation
References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
H2O / DMSO 1 mM 0.9248 mL 4.6242 mL 9.2484 mL 23.1210 mL
5 mM 0.1850 mL 0.9248 mL 1.8497 mL 4.6242 mL
10 mM 0.0925 mL 0.4624 mL 0.9248 mL 2.3121 mL
15 mM 0.0617 mL 0.3083 mL 0.6166 mL 1.5414 mL
20 mM 0.0462 mL 0.2312 mL 0.4624 mL 1.1560 mL
25 mM 0.0370 mL 0.1850 mL 0.3699 mL 0.9248 mL
30 mM 0.0308 mL 0.1541 mL 0.3083 mL 0.7707 mL
40 mM 0.0231 mL 0.1156 mL 0.2312 mL 0.5780 mL
DMSO 50 mM 0.0185 mL 0.0925 mL 0.1850 mL 0.4624 mL
60 mM 0.0154 mL 0.0771 mL 0.1541 mL 0.3853 mL
80 mM 0.0116 mL 0.0578 mL 0.1156 mL 0.2890 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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Neuropeptide FF Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Neuropeptide FF99566-27-5NPFFNeuropeptide FF Receptorrat spinal cordERKNF-κB pathwayadenylate cyclaseG-proteinsacid sensing ion channelGi/o proteinsNPFF receptorhuman NPFF2 receptorMAPK cascadeInhibitorinhibitorinhibit

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