Protamine sulfates, CAS No.9009-65-8

CAS: 9009-65-8 Cat. No.: P123670 EG-Nummer: 618-450-7

Übersicht

This product is provided as delivered and specified by the issuing Pharmacopoeia. All information provided in support of this product, including SDS and any product information leaflets have been developed and issued under the Authority of the issuing Pharmacopoeia.

Specifications

Product Name

Protamine sulfates, CAS No.9009-65-8

Synonyme

Protamines sulfates

CAS

9009-65-8

Molekültyp

Protein

Lagerung und Versand

Storage

Argon charged,Room temperature

Verschickt in

Normal

Application Protocols

No validated application protocols are provided in the Product Data for this item. The following are general literature/practice examples; adjust to your system and consult primary references.

• Example 1: Preparation of protamine working solution

  • Dissolve at 10 mg/mL in sterile water or PBS. Filter through 0.22 µm. Store aliquots refrigerated for short term.

• Example 2: Nucleic acid compaction/precipitation (analytical scale)

  • Mix nucleic acid solution (10–100 µg/mL in low‑salt buffer) with protamine added dropwise to reach N/P 2–5 while gently vortexing.
  • Incubate 15 min at RT; monitor turbidity or run an agarose gel to assess mobility shift/retardation.
  • To reverse, increase NaCl to 0.5–1.0 M and/or add polyanionic competitor.

• Example 3: Heparin binding assay (in vitro)

  • Prepare serial dilutions of protamine (0–200 µg/mL) in assay buffer.
  • Add fixed heparin concentration and incubate 10–20 min; quantify residual free heparin via dye‑binding (e.g., Azure A) or chromatographic readout.

• Example 4: Layer‑by‑layer film assembly

  • Alternate 5–10 min immersions of substrate in protamine (0.5–1 mg/mL, 50–150 mM NaCl) and polyanion solutions, rinsing between steps; track film growth by QCM‑D or UV.

These are starting points only; perform small‑scale optimization for concentration, ionic strength, and timing.

Biological Roles

General/literature biology (no clinical claims):

• Natural function: Protamines are small, arginine‑rich nuclear proteins that replace histones during late spermiogenesis, enabling extreme condensation and stabilization of sperm DNA. The high guanidinium content neutralizes DNA phosphates, and cysteine residues form disulfide crosslinks post‑fertilization.
• Structural features: Predominance of arginine residues yields strong electrostatic and π–cation interactions with nucleic acids; motifs can include serine/cysteine that modulate redox crosslinking and phosphorylation status.
• Conservation and diversity: Species‑specific sequences give rise to multiple protamine isoforms; functional theme is conserved—efficient DNA packaging.
• In vitro utility arising from biology:
  • DNA/RNA condensation: Mimics natural DNA‑compacting environment, applied to assemble compacted nucleic acid particles.
  • Heparin affinity: Strong interaction with glycosaminoglycans mirrors general polyanion binding, useful in biochemical binding studies and affinity capture.
• Biophysical behavior:
  • High pI and polycationic charge drive phase separation/coacervation with polyanions; ionic strength and valency tune condensate properties (literature).

Note: For this catalog item, no organism/source or isoform composition is specified; consult the CoA/Spec Sheet for any lot‑specific sequence or origin information if required for your experiments.

Buffer Applications

Protamine sulfate is not a classical buffering agent; however, it is routinely prepared and used in aqueous buffers for its polyelectrolyte properties.

• Typical working solutions (literature/practice):
  • 1–20 mg/mL in water, PBS, HEPES, or Tris buffers (pH 6.8–7.8). Higher concentrations are feasible depending on ionic strength and desired viscosity.
  • Filter‑sterilize (0.22 µm) if sterility is required. Avoid phosphate concentrations that promote unwanted complexation with co‑present polyanions.
• pH considerations: The reagent remains cationic across neutral pH ranges; buffering capacity is negligible. Select an external buffer suitable for the biological system or assay.
• Ionic strength tuning: Add 50–150 mM NaCl to moderate electrostatic interactions and reduce nonspecific precipitation when handling complex mixtures (cell lysates, serum proteins).
• Practical recipes (literature examples):
  • DNA precipitation aid: 0.1–1 mg/mL protamine in low‑salt TE or water, added dropwise to nucleic acid solution; adjust salt to 100 mM NaCl to promote reversible compaction.
  • Heparin capture assays: Titrate protamine into heparin‑containing buffer while monitoring turbidity or residual heparin via dye‑binding methods.

Because formulation requirements vary widely with sample composition, pilot titrations are recommended to optimize concentration, salt, and pH for each application.

Green Alternatives

From a green chemistry perspective, protamine sulfate is a naturally derived, water‑borne, cationic polyelectrolyte that can replace synthetic polymers in some applications. Considerations (literature/general):

• Renewable sourcing: Often obtained from fish milt (a byproduct stream), providing valorization of biological waste; recombinant sources also exist. Verify sourcing with the supplier if this is critical for your program.
• Benign solvent system: Functions in water, minimizing use of organic solvents in formulation and processing.
• Biodegradability: Peptidic backbone is enzymatically degradable; contrast with persistent synthetic cationic polymers (e.g., PEI).

Comparison (literature):

• Protamine sulfate vs PEI:
  • Advantages: Natural origin, simpler disposal, typically lower environmental persistence; effective polyanion binding at neutral pH.
  • Tradeoffs: Batch heterogeneity; potentially lower stability under harsh pH/temperature; may be costlier per functional charge.
• Protamine sulfate vs Poly‑L‑lysine (PLL):
  • Advantages: Higher charge density per residue due to guanidinium groups; strong binding at physiologic pH.
  • Tradeoffs: Heterogeneous composition vs defined polymer length options available for PLL.
• Operational green wins:
  • Aqueous processing; room‑temperature operations; reduced need for hazardous crosslinkers if relying on electrostatics.

Caveats: For regulated workflows, ensure bioburden/endotoxin levels and lot documentation meet requirements; natural sourcing may introduce variability requiring additional QC.

Pharmaceutical Uses

No therapeutic or clinical use claims are made for this catalog item. The following notes concern research and manufacturing contexts only (literature/general):

• Excipient/process roles in R&D:
  • Polyanion scavenger: Removes anionic impurities (e.g., nucleic acids, glycosaminoglycans) during bioprocess development or analytical sample preparation.
  • Precipitation/clarification aid: Facilitates flocculation of anionic contaminants in protein preparations or fermentation broths.
  • Affinity component: Serves as a heparin‑binding ligand in in vitro assays or as a capture reagent in analytical workflows.
• Formulation research:
  • Prototype complex coacervate systems for encapsulation of anionic actives (e.g., peptides, oligonucleotides) in preclinical formulation studies.
  • Surface modification: Cationic coating to alter zeta potential of particulates or devices in in vitro studies.
• Regulatory compendia: Protamine sulfate materials may appear in pharmacopeial monographs; however, specification alignment for this research‑use product is not stated here. Verify any pharmacopeial compliance directly via the CoA/Spec Sheet if your workflow requires it.

Reminder: This product is for research use only, not for human or animal therapeutic use.

Physical Properties

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.

General/literature properties for protamine sulfate materials (mixtures):

• Molecular weight distribution: Heterogeneous polypeptides typically in the low‑kDa range (often ca. 3–10 kDa, literature), varies by biological source and manufacturing.
• Charge/pI: Strongly basic; isoelectric point frequently >12 (literature), dominated by guanidinium side chains.
• Solubility: Highly soluble in water and physiological buffers; insoluble or sparingly soluble in most organic solvents (ethanol, acetone, ethers) (literature). Forms clear to slightly opalescent solutions depending on ionic strength and concentration.
• Hygroscopicity: Tends to absorb moisture due to high charge density (literature); handle quickly under dry conditions.
• Viscosity: Concentrated aqueous solutions can be moderately viscous; solution behavior depends strongly on ionic strength and pH (literature).
• pKa considerations: Arginine guanidinium groups have high pKa (≈12.5, literature), remaining protonated across a wide pH range; contributes to strong polyelectrolyte behavior.

Not typically defined for this class (mixtures): boiling point, melting point, density, refractive index, and UV cutoff—values depend on composition. For lot‑specific data (e.g., peptide content, ash, sulfate content), refer to the CoA/Spec Sheet.

Quality and Grades

• Item-specific grade/purity: Not specified for this item; refer to CoA/Spec Sheet.

Guidance on interpreting quality for protamine sulfate (general):

• Source and composition: Protamine sulfate is inherently a mixture; quality is commonly characterized by nitrogen content, sulfate content, ash, bioburden/endotoxin levels (for cell‑based work), and peptide molecular weight distribution (SDS‑PAGE or LC‑MS profiling). Review the lot CoA for these parameters.
• Sequence variability: Because protamine is sourced from fish milt or recombinant expression, batch‑to‑batch variability in peptide distribution can influence binding strength to polyanions and precipitation behavior. For sensitive applications (e.g., nucleic acid complexation), validate lot performance.
• UV background: For spectroscopy or chromatography, low UV background and absence of chromophoric impurities may be desirable; if HPLC profiling is relevant, request chromatographic purity traces.
• Stabilizers: Typically none are added; if any stabilizer, antimicrobial, or anti‑caking agent is used, it should be disclosed on the CoA. Absence/presence of salts other than sulfate impacts ionic strength in formulations.
• End-use alignment:
  • Molecular biology/cell culture: Prefer lots tested for bioburden/endotoxin (not specified here—confirm if required).
  • Materials/nanoparticle assembly: Focus on reproducible MW distribution and charge density.
• Documentation: For specifications not listed here, rely on the CoA/Specification Sheet associated with the shipped lot.

Reaction and Applications

This product functions as a cationic polyelectrolyte/reagent rather than a classical substrate or solvent for organic reactions. Common research applications (literature/practice):

• Polyelectrolyte complexation/coacervation:
  • Rapidly forms complexes with polyanions (e.g., DNA, RNA, heparin, alginate, hyaluronan). Used to modulate viscosity, create coacervates, or precipitate target polyanions from solution.
• Nucleic acid handling:
  • Precipitation or compaction of DNA/RNA for purification steps; enhancement of uptake in certain cell systems when used to coat surfaces or assemble complexes (optimize N/P ratio and ionic strength).
• Heparin binding/neutralization in vitro:
  • Useful in biochemical assays to titrate or capture heparin; calibrates heparin assays or removes heparin from protein preparations.
• Nanomaterials and delivery research:
  • Template for inorganic nanoparticle growth; assembly of protein–polysaccharide nanoparticles; layer‑by‑layer films on negatively charged substrates.
• Chromatography/bioprocessing aids:
  • Flocculant for anionic contaminants; can clarify lysates by precipitating nucleic acids and acidic polysaccharides.

Practical tips (literature):

• Work at controlled ionic strength; start with 10–150 mM NaCl and adjust to tune complexation.
• Define N/P ratio (moles of protonated nitrogen to phosphate/sulfate groups) when complexing nucleic acids; typical exploratory range 1–10.
• Add protamine slowly to the polyanion under gentle mixing to avoid local over‑concentration and irreversible aggregation.
• Filter or centrifuge to remove formed complexes; quantify residual polyanion spectroscopically as needed.

Reaction Conditions

Because protamine sulfate acts through electrostatic interactions rather than covalent chemistry, “reaction conditions” refer to assembly/complexation parameters (literature guidance):

• Complexation with polyanions (DNA, heparin, alginate):
  • Solvent: Water or low‑to‑moderate ionic strength buffers (e.g., 10–50 mM HEPES/Tris, 0–150 mM NaCl).
  • pH: 6.5–8.0 maintains peptide integrity and high cationic charge.
  • Stoichiometry: Titrate to target N/P ratio 1–10; monitor turbidity, ζ‑potential, or gel shift to find optimal conditions.
  • Temperature: Ambient (20–25 °C) is typical; avoid elevated temperatures that may promote peptide degradation/aggregation.
  • Mixing: Add protamine slowly to the polyanion under gentle vortexing or stirring; allow 10–30 min equilibration before downstream steps.
• Disassembly/reversibility:
  • Increase ionic strength (e.g., 0.5–1.0 M NaCl) or raise pH moderately to weaken electrostatics; incorporate competitive anions (e.g., polyanionic dyes) if needed.
• Covalent modification (labeling/conjugation):
  • NHS ester labeling in aqueous bicarbonate (pH 8.3) for 30–60 min at RT; quench with Tris or ethanolamine. Degree of labeling controlled by stoichiometry.
• Expected outcomes:
  • Nucleic acid compaction often yields nanoparticles 50–300 nm (literature, system‑dependent) with positive ζ‑potential at N/P >1; yields and sizes are strongly influenced by buffer and mixing.

Note: These are general literature guidelines. Optimize empirically for your specific substrate, buffer composition, and scale.

Safety and Handling

• GHS classification, signal word, pictograms, H‑statements: Not specified for this item; refer to the SDS for authoritative hazard information.
• General hazards (literature/practical):
  • May cause respiratory or skin sensitization in susceptible individuals; peptide/proteinaceous dusts can elicit allergic responses.
  • Irritant potential to eyes, skin, and respiratory tract as a basic polyelectrolyte; can cause local irritation upon contact.
• Special risks/compatibilities:
  • Strongly cationic; will complex and precipitate polyanions (e.g., heparin, nucleic acids, DNA/RNA, glycosaminoglycans). Avoid unintended contact with polyanionic reagents, media supplements, or chromatographic resins unless complexation is desired.
  • Incompatible with strong oxidizers and strong bases/acids that may degrade peptides; avoid prolonged exposure to elevated temperatures and moisture.
• Recommended PPE: Laboratory coat, nitrile gloves, safety glasses; use a dust mask/respirator if handling powders that may become airborne. Work in a fume hood or ventilated enclosure when weighing powders.
• First‑aid overview (consult SDS):
  • Inhalation: Move to fresh air; seek medical attention if symptoms persist.
  • Skin contact: Rinse with water; remove contaminated clothing.
  • Eye contact: Rinse cautiously with water for several minutes; seek medical attention if irritation continues.
  • Ingestion: Rinse mouth; do not induce vomiting; seek medical advice.
• Waste: Dispose according to institutional and local regulations for peptide/protein waste; avoid discharge to drains due to high nitrogen content.
• Storage conditions (from Product Data): Room temperature, Argon charged. Keep container tightly closed to minimize moisture uptake; protect from light and humidity.

Solvent Selection

As a cationic polypeptide salt, protamine sulfate is used primarily in aqueous media rather than as a small‑molecule solute for organic synthesis.

• Primary solvent: Water (ultrapure), physiological buffers (e.g., PBS, HEPES, Tris) at neutral to slightly basic pH. High solubility is typical (literature), though ionic strength modulates complexation.
• Polarity/charge: Strongly cationic across pH 5–10 (literature), favoring interactions with anionic biomolecules; buffer choice strongly affects binding kinetics and precipitation.
• Co‑solvents: Avoid alcohols and high concentrations of kosmotropes when assembling complexes, as these can promote aggregation or precipitation. Low levels of glycerol can stabilize some formulations (literature).
• pH dependence: Because arginine guanidinium groups stay protonated up to high pH, solubility remains robust from pH ~3–10 (literature). Very high pH can promote peptide backbone hydrolysis over time.
• Ionic strength effects: Increasing NaCl or buffer ionic strength screens electrostatic interactions, reducing binding to polyanions and mitigating precipitation. Tailor salt concentration to modulate complex/coacervate formation.
• Comparators (literature):
  • Poly‑L‑lysine (PLL): Similar cationic behavior; lower charge density than protamine at neutral pH.
  • Polyethylenimine (PEI): Stronger polyelectrolyte; branched structure; may exhibit higher cytotoxicity in cell‑based contexts.
  • Chitosan (protonated): Biobased cationic polymer; solubility limited to acidic pH.

Storage and Reconstitution

• Storage conditions (from Product Data): Room temperature, Argon charged. Keep tightly closed in a dry environment to minimize moisture uptake. Store under inert gas as provided to limit oxidation of susceptible residues (e.g., cysteine) and to deter microbial growth.
• Shipping: Normal (from Product Data).

Reconstitution and handling (general/literature guidance):

• Solubility: Readily soluble in water and common biological buffers. Prepare stock solutions at 1–20 mg/mL depending on application.
• Procedure:
  1. Allow container to equilibrate to room temperature before opening to avoid condensation.
  2. Weigh quickly to minimize atmospheric moisture uptake.
  3. Dissolve in desired buffer with gentle stirring; avoid vigorous foaming.
  4. If sterility is required, filter through 0.22 µm PES/RC membrane.
• Solution storage (guidance): For best integrity, store aqueous stocks at 2–8 °C for short term (days to weeks) or aliquot and freeze (−20 °C) for longer term; avoid repeated freeze–thaw. Inspect for turbidity/precipitation before use. These solution recommendations do not override the solid storage instructions above.
• Stability notes: Exposure to high pH, elevated temperatures, or oxidants may promote peptide backbone hydrolysis or disulfide scrambling (literature). Maintain neutral pH and moderate ionic strength unless application requires otherwise.

Always refer to the CoA/Spec Sheet and SDS for lot‑specific guidance on storage, reconstitution, and stability.

Structure and Identity

Protamine sulfates are heterogeneous, arginine‑rich basic polypeptides present as sulfate salts; widely used in biochemical research as strong polycations that bind polyanions (e.g., nucleic acids, glycosaminoglycans).

• Item-specific identifiers (from Product Data):

  • SKU: P123670
  • Product Name: Protamine sulfates
  • CAS: 9009-65-8
  • Molecular Formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.
  • SMILES / InChIKey: Not specified for this item; refer to CoA/Spec Sheet.

• Structural features (general/literature description):

  • Composition: Mixture of low–molecular weight polypeptides highly enriched in L‑arginine and often containing cysteine residues; supplied as sulfate counter‑ions to balance cationic side chains.
  • Functional groups: Guanidinium (from arginine side chains), amide bonds (peptide backbone), occasional disulfides (inter/intramolecular) depending on source and processing.
  • Architecture: Linear peptides; no defined stereocenters beyond the natural L‑amino acid configuration along the backbone; no defined single sequence due to mixture nature.
  • 2D description in words: Repeating peptide backbone (–NH–CH(R)–CO–) with frequent arginine residues presenting guanidinium cations; sulfate serves as inorganic counter‑anion distributed ionically around the polycation.

• Category Path: 全部 / 可售 / 蛋白质 (Protein category)

Note: As a natural‑product–derived mixture, single-molecule descriptors (canonical SMILES, InChI) and exact molecular formula are not typically assigned; consult the CoA for lot‑specific characterization.

Synthetic Utility

While not a classical small‑molecule building block, protamine sulfate has utility in synthetic and materials chemistry (literature):

• Electrostatic templating:
  • Acts as a cationic scaffold to direct the assembly of polyanionic species (e.g., nucleic acids, polyanions, inorganic clusters), enabling formation of nanoparticles, films, and coacervates.
• Catalysis and immobilization supports:
  • Dense guanidinium groups can stabilize transition states for anion‑rich transformations in water (conceptually akin to organocatalytic guanidinium motifs). Protamine‑modified surfaces can immobilize enzymes or anionic catalysts via electrostatics.
• Conjugation chemistry:
  • Primary amines at N‑termini/lysine residues allow covalent modification using NHS esters, isothiocyanates, aldehyde/NaCNBH3 (reductive amination), or click handles introduced through bifunctional linkers.
• Retrosynthetic role in separations:
  • As a precipitation agent, it simplifies downstream purification by removing polyanionic impurities, effectively enabling cleaner access to target biomolecules.
• Layer‑by‑layer (LbL) assemblies:
  • Alternating deposition with polyanions (e.g., PSS, alginate) builds nanofilms; film thickness/porosity tuned by ionic strength and deposition time.

Practical considerations:

• Define and report N/P ratios when forming complexes with defined anionic substrates.
• Dialysis or salt back‑extraction can reverse complexation for recovery of bound polyanions.
• Batch variability (sequence/MW distribution) can affect film formation and particle size—benchmark new lots.

Target Specificity

Not applicable to this product class. Protamine sulfates are not antibodies or affinity‑purified binding proteins with defined epitopes. No target specificity data are provided in the Product Data. For intended binding studies (e.g., heparin, nucleic acids), users should determine conditions empirically and consult literature on polyelectrolyte complexation.