TB-500 Thymosin Beta-4

Research Overview

TB-500 serves as a valuable research tool for investigating fundamental mechanisms of tissue repair, cellular migration, and wound healing in laboratory settings. This synthetic peptide represents the active region of thymosin beta-4, a protein originally isolated from thymus gland tissue and subsequently found to be ubiquitously expressed across mammalian cell types. Research applications have expanded to encompass investigations of cardiovascular repair, musculoskeletal healing, and neurological protection across multiple experimental models.

The peptide’s designation as thymosin beta-4 fragment reflects its origin as part of the thymosin family of proteins involved in immune function and tissue repair. Laboratory studies investigate TB-500’s effects on actin polymerization dynamics, cell migration pathways, extracellular matrix remodeling, and protective mechanisms against various forms of cellular injury. Research protocols examine these effects in cell culture systems, tissue explants, and preclinical animal models.

TB-500 research demonstrates the peptide’s remarkable biological activity through its primary mechanism as an actin-sequestering protein. This fundamental property influences numerous downstream cellular processes including migration, proliferation, differentiation, and survival. Studies have documented TB-500’s ability to promote blood vessel formation, modulate inflammation, reduce fibrosis, and enhance tissue regeneration across diverse experimental contexts.

Molecular Characteristics

Complete Specifications:

• CAS Registry Number: 77591-33-4
• Molecular Weight: 4,963.4 Da
• Molecular Formula: C₂₁₂H₃₅₀N₅₆O₇₈S
• Amino Acid Count: 43 amino acids
• PubChem CID: 16132341
• Peptide Classification: Synthetic thymosin beta-4 analog
• Appearance: White to off-white lyophilized powder
• Solubility: Water, bacteriostatic water, phosphate buffered saline

The peptide’s 43-amino acid structure includes a highly conserved actin-binding domain located within the central region of the sequence. This domain (LKKTET sequence) is critical for TB-500’s ability to sequester G-actin and prevent its polymerization into F-actin filaments. The sequence includes both hydrophobic and hydrophilic regions that contribute to its solubility profile and cellular interactions. The presence of a single cysteine residue contributes to potential disulfide bond formation under oxidizing conditions.

Pharmacokinetic Profile in Research Models

TB-500 pharmacokinetic characterization in preclinical research reveals important properties for experimental design:

Absorption and Distribution:

• Multiple administration routes investigated: IV, IM, SC, IP
• Rapid absorption from subcutaneous and intramuscular sites
• Wide tissue distribution observed across organ systems
• Cellular uptake observed in various cell types in vitro

Metabolism and Elimination:

• Plasma half-life: Approximately 10 days in some animal models
• Slower clearance compared to many peptides of similar size
• Potential intracellular accumulation and sustained activity
• Metabolic stability contributes to prolonged experimental window

These pharmacokinetic characteristics inform research protocol design, particularly regarding dosing frequency and duration in experimental models. The extended half-life compared to many peptides allows for less frequent administration in animal studies while maintaining biological activity.

Research Applications

Tissue Repair and Wound Healing Studies

TB-500 serves as a research tool for investigating fundamental wound healing mechanisms. Laboratory studies examine the peptide’s effects on:

• Cellular Migration Research: Investigation of directional cell movement, chemotaxis, and tissue infiltration during repair processes
• Wound Closure Studies: Analysis of re-epithelialization, wound contraction, and healing rate in various injury models
• Angiogenesis Investigation: Examination of blood vessel formation, endothelial cell sprouting, and vascular network development
• Extracellular Matrix Remodeling: Studies on matrix protein deposition, organization, and turnover during healing
• Inflammatory Modulation: Investigation of inflammatory cell recruitment and resolution during tissue repair

Research protocols typically employ scratch assays, transwell migration assays, and in vivo wound healing models to characterize TB-500’s effects on repair mechanisms.

Cardiovascular Research Applications

Substantial research focuses on cardiovascular tissue investigation:

• Cardiac Protection Studies: Investigation of cardioprotective mechanisms in ischemia-reperfusion models
• Cardiomyocyte Survival Research: Studies examining cardiac cell protection against various injury stimuli
• Coronary Angiogenesis: Research on coronary vessel formation and collateral circulation development
• Cardiac Remodeling Studies: Investigation of post-injury cardiac tissue remodeling and fibrosis
• Endothelial Function Research: Analysis of vascular endothelial cell function and integrity

Laboratory protocols investigate TB-500’s effects in cardiac cell cultures, isolated heart preparations, and animal models of cardiac injury including infarction and ischemia models.

Musculoskeletal Research Applications

Laboratory studies investigate TB-500 in musculoskeletal tissue research:

• Tendon and Ligament Research: Studies on connective tissue healing, collagen fiber organization, and biomechanical strength recovery
• Muscle Tissue Studies: Investigation of muscle fiber regeneration, satellite cell activation, and contractile function restoration
• Skeletal Muscle Injury Models: Research on crush injuries, laceration models, and contusion damage
• Ligament Healing Research: Examination of ligament repair processes, collagen alignment, and mechanical properties
• Tendinopathy Models: Studies investigating mechanisms of tendon pathology and repair

Experimental models include tendon injury models, muscle damage protocols, and ligament transection studies, with outcomes measured through histological analysis, immunohistochemistry, and biomechanical testing.

Neurological and Neuroprotection Research

Research applications extend to nervous system investigation:

• Neuronal Survival Studies: Examination of neuronal protection mechanisms against excitotoxicity and oxidative stress
• Axonal Growth Research: Investigation of neurite outgrowth, axonal extension, and neural pathway development
• Spinal Cord Injury Models: Research on spinal cord tissue protection and functional recovery mechanisms
• Traumatic Brain Injury Studies: Examination of neuroprotective pathways in experimental TBI models
• Neural Stem Cell Research: Studies on neural progenitor cell migration, differentiation, and integration

Laboratory protocols investigate TB-500’s effects on neuronal cell cultures, organotypic brain slice cultures, and rodent models of neural injury.

Inflammatory Modulation Research

Emerging research areas include anti-inflammatory pathway investigation:

• Inflammatory Mediator Studies: Research on cytokine and chemokine expression modulation
• Immune Cell Migration: Investigation of immune cell recruitment and tissue infiltration patterns
• Resolution Phase Research: Studies examining inflammation resolution mechanisms and pro-resolving pathways
• Fibrosis Prevention: Research on excessive collagen deposition and scar tissue formation
• Macrophage Polarization: Investigation of macrophage phenotype switching during tissue repair

Research in this area examines TB-500’s effects on inflammatory cell signaling, cytokine production, and resolution pathway activation in various experimental models.

Laboratory Handling and Storage Protocols

Lyophilized Powder Storage:

• Store at -20°C to -80°C in original sealed vial
• Protect from light exposure and moisture
• Desiccated storage environment recommended
• Stability data available for 24+ months at -20°C

Reconstitution Guidelines:

• Reconstitute with sterile water, bacteriostatic water (0.9% benzyl alcohol), or appropriate buffer
• Add solvent slowly down vial side to minimize foaming
• Gentle swirling motion recommended (avoid vigorous shaking)
• Allow complete dissolution before use (typically 2-3 minutes)
• Final pH should be 6.5-7.5 for optimal stability

Reconstituted Solution Storage:

• Short-term storage: 4°C for up to 14 days
• Long-term storage: -20°C in aliquots to avoid freeze-thaw cycles
• Single-use aliquots recommended to maintain peptide integrity
• Avoid repeated freeze-thaw cycles (maximum 2-3 cycles)

Stability Considerations:
TB-500 demonstrates good stability as a lyophilized powder under proper storage conditions. Reconstituted solutions show reasonable stability at refrigerated temperatures but should be aliquoted for long-term storage to prevent degradation from repeated freeze-thaw cycles.

Quality Assurance and Analytical Testing

Each TB-500 batch undergoes comprehensive analytical characterization:

Purity Analysis:

• High-Performance Liquid Chromatography (HPLC): ≥98% purity
• Analytical method: Reversed-phase HPLC with UV detection at 214nm
• Multiple peak integration to ensure accurate purity determination

Structural Verification:

• Electrospray Ionization Mass Spectrometry (ESI-MS): Confirms molecular weight 4,963.4 Da
• Amino acid analysis: Verifies sequence composition
• Peptide content determination: Quantifies actual peptide content by weight

Contaminant Testing:

• Bacterial endotoxin: <5 EU/mg (LAL method)
• Heavy metals: Below detection limits per USP standards
• Residual solvents: TFA and acetonitrile within acceptable limits
• Water content: Karl Fischer titration (<8%)

Documentation:

• Certificate of Analysis (COA) provided with each batch
• Third-party analytical verification available upon request
• Stability data documented for recommended storage conditions
• Batch-specific QC results traceable by lot number

Research Considerations

Experimental Design Factors:

Researchers should consider several factors when designing TB-500 experiments:

1. Concentration Selection: Determine appropriate concentrations based on research objectives and experimental model. Published research reports ranges from nanomolar to micromolar concentrations depending on application.

2. Temporal Considerations: TB-500’s extended half-life in some models allows for less frequent dosing compared to shorter-acting peptides. Consider timing of measurements relative to administration.

3. Route Considerations: Multiple administration routes show efficacy in research models. Subcutaneous and intraperitoneal routes are commonly used in animal studies.

4. Model Selection: Choose appropriate cell culture systems, tissue explants, or animal models based on specific research questions.

5. Control Groups: Include appropriate vehicle controls, positive controls (where applicable), and comparative compounds.

Mechanism Investigation:

TB-500’s mechanisms of action have been extensively investigated. Primary pathways include:

• Actin sequestration and cytoskeletal dynamics regulation
• PINCH-ILK-α-parvin complex interactions
• VEGF pathway modulation and angiogenesis promotion
• NF-κB signaling pathway modulation
• Laminin-332 upregulation and cell migration enhancement

The peptide’s well-characterized actin-binding mechanism provides a foundation for investigating downstream cellular effects.

Compliance and Safety Information

Regulatory Status:
TB-500 is provided as a research chemical for in-vitro laboratory studies and preclinical research only. This product has not been approved by the FDA for human therapeutic use, dietary supplementation, or medical applications.

Intended Use:

• In-vitro cell culture studies
• In-vivo preclinical research in approved animal models
• Laboratory investigation of biological mechanisms
• Academic and institutional research applications

NOT Intended For:

• Human consumption or administration
• Therapeutic treatment or diagnosis
• Dietary supplementation
• Veterinary therapeutic applications without appropriate oversight

Safety Protocols:
Researchers should follow standard laboratory safety practices when handling TB-500:

• Use appropriate personal protective equipment (lab coat, gloves, safety glasses)
• Handle in well-ventilated areas or fume hood
• Follow institutional biosafety guidelines
• Dispose of waste according to local regulations for biological/chemical waste
• Consult material safety data sheet (MSDS) for additional safety information

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