| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Substance P, FAM-labeled TFA specifically targets the neurokinin 1 receptor (NK1R), a G protein-coupled receptor (GPCR) with high affinity for Substance P. NK1R is widely expressed in the central nervous system, immune cells, and peripheral tissues. Upon binding, Substance P activates Gq/11-mediated signaling, leading to phospholipase C activation, IP3 production, and calcium mobilization. This signaling pathway is involved in pain transmission, inflammation, vasodilation, and smooth muscle contraction. The FAM label does not interfere with receptor binding, allowing for live-cell imaging and binding studies.
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| ln Vitro |
In vitro, Substance P, FAM-labeled TFA (1-100 nM) binds specifically to NK1R-expressing cells (e.g., U373 glioblastoma cells, HEK293-NK1R). The fluorescence signal (λex 488 nm, λem 520 nm) can be detected by flow cytometry, confocal microscopy, or fluorescence plate reader. Binding is competed by excess unlabeled Substance P, confirming specificity. The compound can be used to study NK1R internalization, recycling, and trafficking. It is a high-affinity ligand with a Kd in the low nM range. It is not cytotoxic at concentrations used (1-100 nM).
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| ln Vivo |
Substance P, FAM-labeled TFA is used for ex vivo imaging and in vivo imaging of NK1R expression. It can be injected intravenously (IV) or intracerebroventricularly (ICV) into mice to visualize NK1R-expressing tissues. The fluorescent signal can be detected by whole-body imaging or by fluorescence microscopy of harvested tissues. It is not a therapeutic agent but a research tool for studying the distribution and dynamics of NK1R. The compound crosses the blood-brain barrier poorly due to its peptide nature, so central administration is often required for CNS studies.
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| Enzyme Assay |
The binding affinity of Substance P, FAM-labeled TFA to NK1R is determined by saturation binding assays using membranes from NK1R-expressing cells. Membranes (10-50 ug protein) are incubated with varying concentrations (0.1-100 nM) of FAM-labeled Substance P in binding buffer (50 mM Tris-HCl pH 7.4, 5 mM MgCl2, 0.1% BSA, 0.01% bacitracin) at 25degC for 60-90 min. Bound ligand is separated by filtration or by centrifugation. Fluorescence is measured directly (λex 488 nm, λem 520 nm). Non-specific binding is determined in the presence of a 1000-fold excess of unlabeled Substance P. The Kd is calculated by Scatchard analysis (typically 1-5 nM).
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| Cell Assay |
For NK1R internalization studies, NK1R-expressing cells (e.g., U373 cells) are seeded on coverslips in 24-well plates (2×10⁵ cells/well) and cultured overnight. Cells are treated with Substance P, FAM-labeled TFA (10-100 nM) for 5-60 min at 37degC. Cells are washed, fixed with 4% paraformaldehyde, and counterstained with DAPI. Confocal microscopy is used to visualize cell surface and internalized receptor-bound fluorescence. Receptor internalization is quantified by calculating the ratio of intracellular to cell surface fluorescence. For competition binding assays, flow cytometry is used: cells are incubated with 10 nM FAM-labeled SP with or without excess unlabeled SP (1-10 uM), and mean fluorescence intensity (MFI) is measured.
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| Animal Protocol |
In vivo imaging of NK1R expression: Female BALB/c mice (6-8 weeks old, n=5/group) are injected intravenously (tail vein) with Substance P, FAM-labeled TFA (100-500 ug/kg) in PBS. At 30-60 min post-injection, mice are anesthetized, and whole-body fluorescence imaging is performed using an IVIS Spectrum imaging system (λex 488 nm, λem 520 nm). Mice are euthanized, and organs (brain, spinal cord, spleen, lung, kidney, gut) are harvested for ex vivo fluorescence imaging. For CNS studies, the peptide is administered by intracerebroventricular (ICV) injection (1-10 ug). For pharmacokinetic studies, blood is collected at various time points, and plasma fluorescence is measured.
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| ADME/Pharmacokinetics |
No specific PK data for Substance P, FAM-labeled TFA is available. As a peptide (MW 1706 free base), it has a very short plasma half-life (< 15 min) due to rapid proteolytic degradation. The FAM fluorophore is stable. Following IV injection, it is rapidly cleared from the circulation. It does not cross the blood-brain barrier efficiently. For research use, it is stored as a lyophilized powder at -20degC (protected from light), and it is soluble in DMSO and water.
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| Toxicity/Toxicokinetics |
Substance P, FAM-labeled TFA has low toxicity. Standard safety precautions for handling peptides and fluorescent dyes apply: use PPE (gloves, lab coat, safety goggles), work in a fume hood, avoid inhalation and skin contact. The FAM fluorophore is light-sensitive; protect from light. For research use only.
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| References | |
| Additional Infomation |
Substance P, FAM-labeled TFA is a research-grade fluorescently labeled neuropeptide used for studying NK1 receptor binding, internalization, and trafficking. It is not an FDA-approved drug. It is used for live-cell imaging, flow cytometry, and in vivo imaging of NK1R expression. For research use only, not for diagnostic or therapeutic applications. Storage: powder at -20degC for 3 years, 4degC for 2 years; in solvent at -80degC for 6 months, -20degC for 1 month. Protect from light.
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| Molecular Formula |
C84H108N18O19S.XC2HF3O2
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| Molecular Weight |
1705.93 (free base)
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| Related CAS # |
Substance P, FAM-labeled
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| Appearance |
Light yellow to yellow 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) |
DMSO : ≥ 100 mg/mL
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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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