1. Product Overview
Bufalin (CAS 103144-38-3), systematically named 3β,14-dihydroxy-5β-bufa-20,22-dienolide or (3β,5β,14β)-3,14-dihydroxybufa-20,22-dienolide, is the principal bioactive bufadienolide aglycone derived from the traditional Chinese medicine Chan Su (dried venom of Bufo bufo gargarizans or Bufo melanostictus). With the molecular formula C₂₄H₃₄O₄ and a molecular weight of 386.53 g/mol, it appears as a white to off-white crystalline powder. Bufalin is a potent inhibitor of Na⁺/K⁺-ATPase (IC₅₀ ~40–80 nM in cardiac glycosidase-sensitive preparations), which at low concentrations raises intracellular [Na⁺]→[Ca²⁺] via NCX reversal, and at cytotoxic concentrations activates Src/EGFR, MAPK, PI3K/Akt, NF-κB modulation, induces G2/M or G0/G1 arrest, and triggers mitochondria-mediated apoptosis in a wide range of cancer cells including MDR sublines. It also shows anti-angiogenic, anti-metastatic (MMP suppression), and differentiation-inducing effects. This product is supplied exclusively for laboratory research use and is not intended for clinical, diagnostic, veterinary, food, or cosmetic applications.
2. Key Features and Advantages
• Potent Na⁺/K⁺-ATPase inhibitor & cardiac glycoside mimetic: IC₅₀ ~40–80 nM (ouabain-sensitive site); increases [Ca²⁺]i and activates Src/EGFR cross-talk at sub-cytotoxic doses—enables dissection of signal transducer vs pump functions of Na⁺/K⁺-ATPase.
• Multi-pathway antitumor actions: Downregulates p-Akt (Ser473), p-ERK1/2, p-STAT3; upregulates p-p38 MAPK, Bax/Bak, cleaved caspase-3/9; inhibits TOPO II and HIF-1α in hypoxic tumor cells.
• Overcomes MDR: Retains activity in P-gp-overexpressing cells (K562/ADR, MCF-7/ADR) partly via non-Pgp substrate behavior and modulation of MDR1 expression.
• Anti-angiogenesis & anti-metastasis: Inhibits HUVEC tube formation, MMP-2/9 secretion, and VEGF expression; suppresses EMT markers (↓Snail, ↑E-cadherin).
• High purity: Typically ≥98% by HPLC (suppliers offer 98–99%+), suitable for MTT/XTT, Annexin V/PI flow, Western blot (p-Akt, p-ERK, cleaved caspase-3, LC3-II), intracellular Ca²⁺ imaging (Fura-2 AM), and LC-MS quantification from biological matrices.
• Soluble in DMSO (≥5 mg/mL with mild warming), ethanol, methanol; poorly soluble in water; light-sensitive (bufadienolide lactone).
3. Main Applications
• Na⁺/K⁺-ATPase signaling research: Using bufalin as a tool to activate Src/EGFR crosstalk via Na⁺/K⁺-ATPase α-subunit (distinct from ouabain at different conc. ranges); comparing with ouabain, digoxin.
• Antitumor mechanism & MDR studies: Evaluating G2/M or G0/G1 arrest, apoptosis induction, autophagy, and ROS generation in leukemia (HL-60, K562, K562/ADR), liver (HepG2), breast (MCF-7, MDA-MB-231), gastric, and prostate cancer lines.
• Angiogenesis & metastasis: In vitro (HUVEC tube, Transwell invasion, zymography for MMP-2/9) and in vivo (Matrigel plug, zebrafish ISV) models of neo-vascularization and EMT reversal.
• Traditional medicine & natural product research: HPLC quantification in Chan Su / toad venom extracts; SAR of bufadienolide aglycones vs glycosides (bufalin vs bufotalin, cinobufagin).
• Differentiation induction: Testing myeloid differentiation (CD11b, CD14 ↑, CD33 ↓) in HL-60 and U937 promyelocytic leukemia models.
4. Technical Specifications
5. Storage and Handling
• Store the powdered compound sealed with desiccant, strictly protected from light (amber vial or foil-wrapped); recommended storage at -20°C (long term 2 years) or 4°C (≤6 months). For DMSO or ethanolic stock solutions, prepare fresh when possible; if storing, aliquot and keep at -20°C or -80°C wrapped in foil (3–6 months stable); avoid repeated freeze–thaw cycles. Typical working stocks are 5 mM or 10 mM in anhydrous DMSO, prepared with gentle warming (≤40°C) if needed.
• Before opening, allow the sealed vial to warm to room temperature to prevent moisture condensation. Keep container tightly closed and wrapped in aluminum foil when not in use.
• Handle in a well-ventilated area or fume hood. Use personal protective equipment including safety glasses, nitrile gloves, and a lab coat. Avoid inhalation of dust and contact with skin or eyes. Bufadienolides are potent cardiotonic steroids—exercise extreme caution, especially with injection-form or high-concentration stocks.
• Intended exclusively for laboratory research by qualified personnel; not for human or veterinary use, food additive, or TCM preparation (despite being a TCM-derived constituent—this material is research-grade only).
6. Safety Overview
• Bufalin is a potent cardiotonic steroid (cardiac glycoside analog) with cytotoxicity (IC₅₀ 10–200 nM in many cancer lines) and significant in vivo toxicity (arrhythmogenic at low doses in animals). Treat as highly hazardous: may cause severe irritation to eyes, skin, and respiratory system; suspected carcinogen/mutagen; digitoxicity risk.
• In case of eye contact, rinse immediately with plenty of running water for at least 15 minutes and seek immediate medical attention. In case of skin contact, wash thoroughly with soap and water for several minutes.
• If inhaled, move the person to fresh air and keep at rest in a position comfortable for breathing; seek medical advice promptly. If ingested, rinse mouth and seek emergency medical attention—do not induce vomiting.
• Use appropriate engineering controls (fume hood, closed handling) and personal protective equipment (safety goggles, nitrile gloves, double gloving recommended for powder weighing, lab coat, face shield). Avoid generating dust; work under reduced light due to photosensitivity.
• Dispose of in accordance with local regulations for hazardous chemical, cytotoxic, and natural toxin waste. Do not discharge into drains, waterways, or soil.
• For detailed safety information, refer to the applicable Safety Data Sheet (SDS) prior to handling; many institutions require special authorization for bufadienolide/cardiac glycoside work.
