| 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 |
Bak 26 nM (Kd)
PUMA BH3 targets the pro-apoptotic protein Bak (Bcl-2 homologous antagonist/killer). It binds to Bak in the canonical BH3-binding groove, inducing Bak conformational changes, homo-oligomerization, and Bak-mediated membrane permeabilization. PUMA BH3 also binds to anti-apoptotic proteins (Bcl-2, Bcl-xL, Mcl-1) as a BH3-only protein, neutralizing them and liberating pro-apoptotic effectors. The Kd for Bak is 26 nM. |
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| ln Vitro |
At a Kd of 26 nM, PUMA BH3, a peptide containing the BH3 domain and increased by p53, functions as a direct activator of Bak. PUMA BH3 activates Bak by attaching to it in the standard BH3 binding groove[1].
In vitro, PUMA BH3 acts as a direct activator of Bak with a dissociation constant (Kd) of 26 nM (in CHAPS buffer) or 290 nM (in HEPES buffer). It induces Bak homo-oligomerization and Bak-mediated membrane permeabilization. PUMA BH3 binds to Bak in the canonical BH3-binding groove and activates Bak. The peptide is a member of the BH3-only family and is a key mediator of p53-induced apoptosis. |
| ln Vivo |
In vivo, PUMA is a p53-upregulated modulator of apoptosis. PUMA proteins bind Bcl-2, localize to the mitochondria, and induce cytochrome C release and apoptosis in response to p53, potentially mediating p53-induced apoptosis. The BH3 domain peptide derived from PUMA retains this pro-apoptotic function. PUMA BH3 can be used in vivo to study apoptosis regulation and BH3 mimetics.
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| Enzyme Assay |
PUMA BH3 TFA is a peptide, and its binding to Bak is characterized using a cell-free binding assay. Recombinant Bak protein (10-50 nM) is incubated with increasing concentrations (0.1-1000 nM) of PUMA BH3 peptide in binding buffer (HEPES or CHAPS buffer). A fluorescently labeled BH3 peptide tracer (e.g., FITC-Bid BH3) is added. Fluorescence polarization (FP) is measured, and the Kd is calculated from the competition curve. The Kd for Bak is 26 nM in CHAPS buffer.
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| Cell Assay |
PUMA BH3 is used in cell-based apoptosis assays. Cancer cells are seeded in 96-well plates in appropriate medium. PUMA BH3 peptide can be delivered into cells using a cell-penetrating peptide fusion or a transfection reagent. Cells are treated with PUMA BH3 (1-50 uM) for 24-48 hours. Cell viability is measured by MTT or CellTiter-Glo. Apoptosis is assessed by Annexin V-FITC/PI staining and flow cytometry. Cytochrome C release from mitochondria is measured by Western blot. Caspase-3/7 activity is quantified.
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| Animal Protocol |
PUMA BH3 can be used in vivo in mouse tumor xenograft models. Female BALB/c nude mice are implanted subcutaneously with cancer cells. When tumors reach 100-150 mm3, mice are randomized. PUMA BH3 peptide (dissolved in PBS) is administered intratumorally (5-20 ug/mouse) or intravenously (1-5 mg/kg) in a formulation such as 10% DMSO, 40% PEG300, 5% Tween 80, 45% saline. Tumor volume is measured every 2-3 days. Apoptosis in tumor tissues is assessed by TUNEL and cleaved caspase-3 IHC.
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| ADME/Pharmacokinetics |
PUMA BH3 is a peptide with a molecular weight of 3163.32 and the sequence Ac-Glu-Gln-Trp-Ala-Arg-Glu-Ile-Gly-Ala-Gln-Leu-Arg-Arg-Met-Ala-Asp-Asp-Leu-Asn-Ala-NH2. The powder should be stored at -20degC for up to 3 years. In solvent, it is stable for 1 year at -80degC. The TFA salt is the standard commercial form. For in vivo use, formulate in 10% DMSO, 40% PEG300, 5% Tween 80, 45% saline.
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| Toxicity/Toxicokinetics |
PUMA BH3 is a peptide derived from a pro-apoptotic protein; at high concentrations or with prolonged exposure, it may cause non-specific cytotoxicity. In animal models at the doses used, no overt toxicity has been reported. The TFA counterion presents no novel safety concerns at the low doses used. Standard safety precautions for peptide handling should be followed.
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| References | |
| Additional Infomation |
PUMA BH3 TFA is a research-grade peptide and is not approved for clinical use. It is a pro-apoptotic peptide derived from the BH3 domain of PUMA that directly activates Bak with a Kd of 26 nM. It binds to Bak in the canonical BH3-binding groove and activates Bak. This product is for research use only and not for human therapeutic applications.
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| Molecular Formula |
C100H162N34O32S
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| Molecular Weight |
2384.63
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| Related CAS # |
PUMA BH3 TFA
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| Appearance |
White to off-white 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 (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 :~100 mg/mL (~41.94 mM)
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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 | 0.4194 mL | 2.0968 mL | 4.1935 mL | |
| 5 mM | 0.0839 mL | 0.4194 mL | 0.8387 mL | |
| 10 mM | 0.0419 mL | 0.2097 mL | 0.4194 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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