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| Targets |
The primary target of OVA Peptide (257-264) TFA is the MHC class I molecule H-2Kb, which is expressed on the surface of antigen-presenting cells (APCs) in C57BL/6 mice. The peptide binds to the peptide-binding groove of H-2Kb with high affinity, forming a stable peptide-MHC complex that is recognized by the T-cell receptor (TCR) on CD8+ cytotoxic T lymphocytes (CTLs). This interaction is the basis for the peptide's use in studying antigen-specific T-cell responses. Upon binding to H-2Kb, the peptide-MHC complex is displayed on the cell surface, where it can activate CD8+ T cells that express a TCR specific for the SIINFEKL-H-2Kb complex. This leads to T-cell proliferation, cytokine production, and the generation of cytotoxic T-cell responses.
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| ln Vitro |
TAP1-I- and C57BL/6 macrophages can process Crl-OVA and full-length OVA in distinct cellular compartments. Additionally, TAP-independent processing of MHC class I presentation is influenced by the protein background of the OVA epitope, OVA peptide (257-264). The TAP transporter presents the epitope in several ways: While the OVA peptide (257–264) in full-length ovalbumin is highly dependent on the TAP transporter, regardless of whether the native protein is coupled to polystyrene or the recombinant OVA is expressed in bacteria, the OVA peptide (257–264) in the MBPCrl-OVA or Crl-OVA bacterial fusion protein depends on the TAP transporter [1].
The in vitro activity of OVA Peptide (257-264) TFA is defined by its ability to bind to H-2Kb and activate CD8+ T cells. In peptide-MHC binding assays, the peptide demonstrates high affinity for H-2Kb, with an IC₅₀ in the nanomolar range. In T-cell activation assays, the peptide is used to stimulate OVA-specific CD8+ T cells, such as those from OT-I TCR transgenic mice, leading to robust T-cell proliferation and cytokine production (e.g., IFN-γ, IL-2). The peptide's activity is dependent on its presentation by H-2Kb on APCs, and it is commonly used to pulse APCs for T-cell stimulation assays. Its high purity (≥98%) ensures reliable and reproducible results in these assays. |
| ln Vivo |
In vivo, OVA Peptide (257-264) TFA is used to induce and study antigen-specific CD8+ T-cell responses in C57BL/6 mice. When administered via injection (e.g., intraperitoneal, subcutaneous, or intravenous), the peptide is taken up by APCs, processed, and presented on H-2Kb, leading to the activation and expansion of OVA-specific CD8+ T cells. This model is widely used to study the mechanisms of T-cell activation, memory formation, and immunotherapy. The peptide can also be used in combination with adjuvants to enhance the immune response. Its in vivo effects are dependent on the presence of H-2Kb and OVA-specific TCRs.
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| Enzyme Assay |
In vitro peptide-MHC binding assays are performed to measure the affinity of OVA Peptide (257-264) for H-2Kb. In a typical assay, recombinant H-2Kb protein or cell lysates from H-2Kb-expressing cells are incubated with varying concentrations of the peptide and a fluorescently labeled or radiolabeled reference peptide. After incubation, the amount of bound peptide is quantified using methods such as fluorescence polarization, scintillation counting, or flow cytometry. The concentration of the peptide required to displace 50% of the labeled peptide (IC₅₀) is calculated. T-cell activation assays are performed by pulsing APCs with the peptide and co-culturing them with OVA-specific CD8+ T cells, followed by measurement of T-cell proliferation (e.g., by [³H]-thymidine incorporation or CFSE dilution) and cytokine production (e.g., by ELISA or ELISpot).
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| Cell Assay |
In vitro cell-based assays for OVA Peptide (257-264) TFA are performed using antigen-presenting cells (APCs) and T cells. APCs (e.g., bone marrow-derived dendritic cells, splenocytes, or cell lines such as RMA-S or EL4) are pulsed with the peptide for 1-4 hours, washed, and then co-cultured with OVA-specific CD8+ T cells (e.g., from OT-I mice). T-cell activation is measured by assessing proliferation (via [³H]-thymidine incorporation, CFSE dilution, or CellTrace Violet), cytokine production (IFN-γ, TNF-α, IL-2 by ELISA or intracellular cytokine staining), and upregulation of activation markers (CD69, CD25, CD44) by flow cytometry. These assays are fundamental for studying T-cell biology and for evaluating the immunogenicity of peptides and vaccines.
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| Animal Protocol |
In vivo animal studies for OVA Peptide (257-264) TFA are typically conducted in C57BL/6 mice. A common protocol involves immunizing mice with the peptide, often in combination with an adjuvant (e.g., complete Freund's adjuvant, poly(I:C), or CpG), via subcutaneous, intraperitoneal, or intravenous injection. The immune response is assessed by measuring the frequency of OVA-specific CD8+ T cells in the blood, spleen, and lymph nodes using MHC class I tetramers (H-2Kb/SIINFEKL) or by intracellular cytokine staining after peptide restimulation. The peptide can also be used in adoptive transfer studies, where OVA-specific T cells (e.g., from OT-I mice) are transferred into recipient mice, which are then immunized with the peptide to study T-cell dynamics and function.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of OVA Peptide (257-264) TFA are characteristic of a peptide. As a peptide, it is susceptible to rapid proteolytic degradation by peptidases in the blood and tissues, which results in a very short half-life. It is not expected to be orally bioavailable and is typically administered via injection. Its volume of distribution is likely limited to the extracellular space. Clearance occurs primarily through metabolism. For research purposes, the peptide is typically dissolved in sterile water, PBS, or saline. The TFA salt form is used to enhance the peptide's solubility and stability. The peptide should be stored as a lyophilized powder at -20°C to maintain its integrity.
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| Toxicity/Toxicokinetics |
The toxicological profile of OVA Peptide (257-264) TFA is not extensively documented, as it is a research tool rather than a therapeutic agent. At the concentrations used in immunological assays (typically in the μg/mL to mg/mL range), it is not expected to be toxic. In vivo, the peptide is generally well-tolerated, and adverse effects are not typically observed at immunogenic doses. However, the use of adjuvants can cause local inflammation and other side effects. For laboratory handling, standard safety precautions for research chemicals should be observed, including the use of personal protective equipment (gloves, lab coat, safety goggles). The compound is intended for research use only and is not for human therapeutic or diagnostic use.
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| References | |
| Additional Infomation |
OVA Peptide (257-264) TFA is a widely used research tool in immunology. It is the class I (Kb)-restricted peptide epitope of ovalbumin, with the sequence SIINFEKL. It is presented by the MHC class I molecule H-2Kb in C57BL/6 mice. The peptide is used in a multitude of applications, including T-cell activation assays, tetramer staining, and in vivo immunization studies. It has a molecular formula of C₄₉H₇₆F₆N₁₀O₁₇ and a molecular weight of approximately 1191.17 g/mol. It is supplied as a white to off-white solid powder with a purity of ≥98%. OVA Peptide (257-264) TFA is for research use only.
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| Molecular Formula |
C49H76F6N10O17
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|---|---|
| Molecular Weight |
1191.17497444153
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| Exact Mass |
1076.536
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| CAS # |
1262751-08-5
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| Related CAS # |
OVA Peptide(257-264);138831-86-4;OVA Peptide(257-264) acetate salt
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| PubChem CID |
71311993
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
14
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| Hydrogen Bond Acceptor Count |
20
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| Rotatable Bond Count |
33
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| Heavy Atom Count |
75
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| Complexity |
1780
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| Defined Atom Stereocenter Count |
10
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| SMILES |
FC(C(=O)O)(F)F.FC(C(=O)O)(F)F.O=C([C@H]([C@@H](C)CC)NC([C@H](CO)N)=O)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)CC(C)C)=O)CCCCN)=O)CCC(=O)O)=O)CC1C=CC=CC=1)=O)CC(N)=O)=O)[C@@H](C)CC
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| InChi Key |
BRUCWCULSHVYBQ-VPZPQSBKSA-N
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| InChi Code |
InChI=1S/C45H74N10O13.C2HF3O2/c1-7-25(5)36(55-44(66)37(26(6)8-2)54-38(60)28(47)23-56)43(65)52-32(22-34(48)57)42(64)51-31(21-27-14-10-9-11-15-27)41(63)50-30(17-18-35(58)59)40(62)49-29(16-12-13-19-46)39(61)53-33(45(67)68)20-24(3)4;3-2(4,5)1(6)7/h9-11,14-15,24-26,28-33,36-37,56H,7-8,12-13,16-23,46-47H2,1-6H3,(H2,48,57)(H,49,62)(H,50,63)(H,51,64)(H,52,65)(H,53,61)(H,54,60)(H,55,66)(H,58,59)(H,67,68);(H,6,7)/t25-,26-,28-,29-,30-,31-,32-,33-,36-,37-;/m0./s1
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| Chemical Name |
(2S)-2-[[(2S)-6-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S,3S)-2-[[(2S,3S)-2-[[(2S)-2-amino-3-hydroxypropanoyl]amino]-3-methylpentanoyl]amino]-3-methylpentanoyl]amino]-4-oxobutanoyl]amino]-3-phenylpropanoyl]amino]-4-carboxybutanoyl]amino]hexanoyl]amino]-4-methylpentanoic acid;2,2,2-trifluoroacetic 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 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 : ~125 mg/mL (~116.05 mM)
H2O : ~50 mg/mL (~46.42 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (1.93 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.8395 mL | 4.1975 mL | 8.3950 mL | |
| 5 mM | 0.1679 mL | 0.8395 mL | 1.6790 mL | |
| 10 mM | 0.0840 mL | 0.4198 mL | 0.8395 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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