BS2G Crosslinker-ddisodium , CAS No.910292-86-3

CAS: 910292-86-3 Cat. No.: B1440530 Summenformel: C13H10D4N2O8 Molekulargewicht: 330.28

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Übersicht

BS2G Crosslinker-d 4 (disodium) is the deuterium labeled BS2G Crosslinker. Radioactive isotopes labeled compounds can be used in metabolic analysis, allowing the movement of individual atoms to be precisely tracked and quantified.

Specifications

Storage

Store at -20°C

Verschickt in

Ice chest + Ice pads

Dieses Produkt erfordert Kühlkettenversand. Grundversand und andere Economy-Optionen sind nicht verfügbar.

Namen und Kennungen

Molekulargewicht

330.28

Application Protocols

No item‑specific, validated protocols are provided in the Product Data. The following is a general, literature‑based workflow for protein crosslinking with BS2G.

General protein–protein crosslinking (literature/general):

1. Buffer exchange: Place proteins in PBS or HEPES (pH 7.2–7.8), 0.1–1 mg/mL. Remove amine buffers (Tris/glycine) by desalting.
2. Prepare crosslinker: Dissolve BS2G freshly at 5–20 mM in water or buffer (or 50–200 mM in DMSO/DMF, then spike into reaction to 0.25–5 mM final). Keep cold, use within minutes.
3. Reaction: Add BS2G to protein while gently mixing. Incubate 10–45 min at 0–25°C. For XL‑MS, shorter times and lower equivalents help control crosslink density.
4. Quench: Add 20–50 mM Tris or glycine; incubate 10–20 min.
5. Analysis: Evaluate by SDS‑PAGE (mobility shifts), SEC (oligomerization), or MS (identify crosslinked peptides). For immobilization, wash surfaces thoroughly and block remaining sites with ethanolamine or Tris.

Surface coupling (literature/general):

• Clean amine‑functional surfaces, equilibrate in buffer, apply 1–10 mM BS2G for 10–30 min, rinse, then quench. Optimize time and concentration to balance surface coverage and nonspecific adsorption.

Note: Adjust conditions to biomolecule stability. Item‑specific instructions are Not specified for this item; refer to CoA/Spec Sheet.

Biological Roles

Not a natural metabolite; BS2G is a synthetic crosslinking reagent for laboratory use. It does not have intrinsic biological roles.

General/literature context:

• Mode of action in bioconjugation: Chemically reacts with primary amines on biomolecules to form amide bonds; reactivity is purely chemical, not enzymatic or receptor‑mediated.
• Cell permeability: The sulfonate groups render the reagent membrane‑impermeant, confining reactions to extracellular or surface‑accessible amines in cell‑based workflows (literature/general).
• Impact on biomolecules: Crosslinking can stabilize or immobilize proteins and complexes; excessive modification may perturb function. Titrate carefully to balance coupling efficiency and biological activity.

No medical or clinical claims are made. For research use only (from Product Data).

Buffer Applications

Highly applicable. Successful use of BS2G depends strongly on buffer composition and pH.

Recommended systems (literature/general):

• Compatible (no primary amines):
  • Phosphate buffers (PBS, 10–100 mM, pH 7.2–7.8) for general protein conjugation.
  • HEPES or MOPS (25–50 mM, pH 7.2–7.8) when phosphate interactions are undesirable.
  • Bicarbonate/carbonate (50–100 mM, pH 8.3–8.5) for faster reactions with robust targets.
• Incompatible (consume reagent):
  • Tris, glycine, ethanolamine, or other primary/secondary amine buffers; avoid until quenching.

Practical buffer tips:

• Degas buffers gently if working oxygen‑sensitive systems; filter (0.22 µm) to remove particulates that may present nucleophilic sites.
• Include salts (e.g., 150 mM NaCl) to maintain protein stability; avoid chelators only if metal‑dependent proteins are under study.
• For hydrophobic substrates, use ≤10–20% DMSO/DMF cosolvent; verify biomolecule tolerance.

Quenching (literature/general):

• After reaching desired coupling, add 20–100 mM Tris or glycine to cap residual esters; incubate 10–30 min at reaction temperature.

Item-specific buffer recipes or validated conditions: Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

Perspective: “Greener” in this context relates to solvent choices, atom economy, and hazard reduction during bioconjugation. While BS2G is itself a specialized reagent, you can improve sustainability around its use.

Greener choices (literature/general):

• Solvent/buffer system:
  • Prefer aqueous buffers (phosphate, HEPES) at ambient temperature over high‑percent organic media. Limit DMF/DMSO to the minimum needed for solubilization.
  • Use bicarbonate/carbonate buffers prepared from CO2‑neutral sources where feasible; avoid amine buffers to prevent consumption of reagent.
• Alternative coupling chemistries:
  • Zero‑length coupling (EDC with NHS or sulfo‑NHS) can be more atom‑economical when linking carboxylates to amines, reducing persistent spacer fragments but with different selectivity.
  • Enzymatic ligations (e.g., sortase A, microbial transglutaminase) provide high selectivity in water but require engineered motifs.
• Process improvements:
  • Miniaturize reactions, use single‑use aliquots to reduce waste from hydrolyzed solutions.
  • Quench residual activity and neutralize solutions before disposal to minimize ecotoxicity concerns.

Tradeoffs:

• BS2G offers rapid, broad amine reactivity in water, but hydrolysis leads to succinimide byproducts and requires stoichiometric reagent. Enzymatic or click‑type strategies can lower waste but need specific handles and may not be suitable for native systems.

Item-specific environmental data (biodegradability, VOC content, etc.): Not specified for this item; refer to SDS.

Pharmaceutical Uses

This product is supplied for research use only (from Product Data). It is not intended for human or veterinary use.

General, non‑clinical context (literature/general):

• Process reagent: Sulfo‑NHS ester crosslinkers like BS2G may be used during R&D for preparing prototype conjugates, immobilized enzymes, or analytical tools. Such uses are confined to preclinical research settings.
• Excipient status: No pharmacopeial excipient status is implied. Any cGMP or clinical manufacturing use would require dedicated qualification and is outside the scope of this listing.

Item‑specific regulatory designations, residuals limits, or compendial compliance: Not specified for this item; refer to CoA/Spec Sheet.

Physical Properties

Only limited item-specific physical data are provided. Do not construe the following literature notes as specifications.

Item-specific (from Product Data):

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.

General/literature characteristics for sulfo‑NHS glutarate crosslinkers:

• Physical form: typically off‑white to white, hygroscopic solid (literature/general).
• Solubility: generally readily soluble in aqueous buffers (pH ~7–8.5); also soluble in polar organics such as DMF/DMSO (literature/general).
• Partitioning: hydrophilic due to sulfonate groups; effectively membrane‑impermeant (literature/general).
• Hydrolytic lability: activated esters hydrolyze in water; rate increases with pH, temperature, and buffer nucleophilicity (literature/general).
• Stability: enhanced when dry, cold, and protected from moisture; solutions are short‑lived (literature/general).

Commonly cited handling ranges (literature/general, not product specifications):

• Working concentration: 0.1–10 mM in aqueous buffer for protein crosslinking, prepared immediately before use.
• pH window for amine coupling: ~7.0–8.5 (balance between amine nucleophilicity and ester hydrolysis).

Any exact values such as melting point, water content, UV cutoff, or density are Not specified for this item; refer to CoA/Spec Sheet and SDS for authoritative information.

Quality and Grades

Item-specific grade/purity is not listed in the Product Data; consult the CoA/Spec Sheet for measured values and acceptance criteria.

Context for this reagent type (literature/general):

• Crosslinker/research grade: Typically indicates suitability for biochemical conjugations, with characterization by NMR/HRMS/HPLC to ensure identity and low levels of hydrolysis byproducts. Low residual solvents and controlled inorganic content reduce assay interference.
• Moisture control: Because NHS esters hydrolyze, high-quality lots emphasize low water content and packaging under dry inert conditions. Single‑use or small‑volume vials help maintain activity.
• Stabilizers: Sulfo‑NHS crosslinkers are generally supplied neat without stabilizers to avoid introducing nucleophiles. If any stabilizer or desiccant system is present, it will be stated on the product label/CoA (Not specified for this item; refer to CoA/Spec Sheet).
• QC tests you may expect (literature/general): appearance, assay by HPLC, identity by NMR/HRMS, residual solvents (GC), water (KF), and functional performance tests (model amine coupling). Exact specifications are Not specified for this item; refer to CoA/Spec Sheet.

Practical note:

• For sensitive bioconjugations (e.g., quantitative interfacial crosslinking), request the latest CoA with purity, water content, and recommended handling window. Verify lot‑to‑lot performance by a small‑scale pilot conjugation.

Reaction and Applications

Relevance: Central to this reagent. The following guidance is derived from general literature on sulfo‑NHS amine‑reactive crosslinkers.

Primary reaction class:

• N‑acylation of primary amines via activated sulfo‑NHS esters to form stable amide bonds. BS2G is homobifunctional, enabling intra‑ and intermolecular crosslinks between lysine side chains or N‑termini on proteins/peptides and amine‑functionalized surfaces.

Common applications (literature/general):

• Protein–protein crosslinking to probe complex assembly, stabilize quaternary structures, or capture transient interactions for MS mapping.
• Surface bioconjugation on amine‑bearing polymers, beads, hydrogels, and nanoparticles where water solubility and membrane impermeance are beneficial.
• Antibody immobilization and orientation control via controlled amine coupling (avoiding antigen‑binding site when possible by pH/stoichiometry management).
• Crosslinking of cells/tissues at the extracellular surface for receptor clustering studies (membrane‑impermeant profile).

Practical tips:

• Prepare solutions immediately before use (e.g., 5–20 mM stocks in water or PBS, or 50–200 mM in DMF/DMSO for spiking) to limit hydrolysis. Keep on ice just prior to addition.
• Control reaction by stoichiometry and time: excess BS2G increases crosslink density; quench with Tris, ethanolamine, or glycine after desired coupling time to cap remaining esters.
• Temperature: perform at 0–25°C depending on biomolecule stability; lower temperatures reduce hydrolysis and preserve activity (literature/general).
• Analysis: Monitor consumption by UV/HPLC or infer from functional readouts; validate crosslinks by SDS‑PAGE shift, SEC, or MS crosslink mapping.

Note: Item‑specific recommended conditions are Not specified for this item; refer to CoA/Spec Sheet and SDS.

Reaction Conditions

The following are general, literature‑based guidelines for sulfo‑NHS glutarate crosslinkers; they are not product specifications.

Typical conditions (literature/general):

• Concentration: 0.25–5 mM BS2G vs. 0.1–10 mg/mL protein. For surfaces, adjust to surface amine density; 1–10 mM is common for dense monolayers.
• Solvent/buffer: PBS, HEPES, or bicarbonate (pH 7.2–8.5). Avoid amine buffers. Optional 5–10% DMSO/DMF for solubility.
• Temperature/time: 0–25°C for 10–60 minutes. Lower temperature reduces hydrolysis for sensitive proteins; higher pH/temperature shortens required time but increases side hydrolysis.
• Stoichiometry: 5–50 equivalents of BS2G per accessible amine for proteins; titrate to modulate crosslink density.
• Quench: Add 20–100 mM Tris or glycine; incubate 10–30 min.

Operational tips:

• Prepare fresh solutions immediately before use; keep on ice and add last to the reaction to minimize premature hydrolysis.
• For XL‑MS, short reaction times (e.g., 2–10 min) and lower equivalents can reduce over‑crosslinking and preserve complex topology.
• For surfaces, pre‑equilibrate substrate in buffer, remove excess water, and apply BS2G solution uniformly; rinse after coupling and quench.

Expected outcomes (literature/general):

• Efficient amide bond formation with primary amines; minor hydrolysis byproducts (N‑hydroxysulfosuccinimide and glutarate monoesters/diacids). Yields depend on substrate accessibility and reaction control.

Item‑specific validated conditions: Not specified for this item; refer to CoA/Spec Sheet.

Safety and Handling

Authoritative safety information resides in the SDS. The following are prudent, general practices for sulfo‑NHS ester crosslinkers.

Item-specific hazard data (from Product Data):

• Signal word: Not specified for this item; refer to SDS.
• H‑Statements: Not specified for this item; refer to SDS.
• GHS classification/pictograms: Not specified for this item; refer to SDS.

General precautions (literature/general):

• Handle in a fume hood or well‑ventilated area; avoid inhalation of dust and contact with skin/eyes. Wear appropriate PPE: lab coat, safety glasses/goggles, and chemically resistant gloves (nitrile). Avoid moisture exposure—product is moisture‑sensitive and can hydrolyze to byproducts.
• Incompatibilities: water, alcohols, amine‑containing buffers (e.g., Tris, glycine) during storage/use; strong bases/acids accelerate hydrolysis; reducing agents and nucleophiles may consume the reagent.
• First aid (overview; see SDS): in case of skin/eye contact, rinse with copious water for at least 15 minutes and seek medical attention. If inhaled, move to fresh air. If ingested, rinse mouth; do not induce vomiting unless directed by medical personnel.
• Waste: Collect aqueous/organic waste containing activated esters in labeled containers; quench residual activity with excess primary amine or hydrolyze under controlled basic conditions before disposal in accordance with institutional and local regulations.
• Special risks: Hydrolysis liberates N‑hydroxysulfosuccinimide; avoid accumulation of dry dusts. Do not scale up without risk assessment.

Solvent Selection

Applicability: Highly relevant. BS2G (sulfo‑NHS glutarate, disodium) is generally chosen for aqueous, membrane‑impermeant crosslinking.

General/literature guidance:

• Polarity and miscibility: Sulfonate groups confer high water solubility; BS2G dissolves readily in neutral to slightly basic buffers (pH ~7–8.5). It is also soluble in polar aprotics (DMF, DMSO) and can be introduced as a concentrated stock into aqueous media.
• Buffer compatibility: Avoid primary amine buffers (Tris, glycine, ethanolamine) that consume the reagent. Preferred: phosphate (PBS), HEPES, MOPS, bicarbonate/carbonate. Maintain ionic strength compatible with your biomolecules.
• pH considerations: Higher pH increases amine nucleophilicity but accelerates hydrolysis; pH 7.2–7.8 is a common compromise for proteins. For rapid surface derivatization on robust substrates, pH 8.3–8.5 may be used with shortened reaction times (literature/general).
• Cosolvents: For hydrophobic samples, up to 10–20% DMF or DMSO can aid solubilization without unduly quenching activity; verify biomolecule stability. Add organic cosolvent to the sample first, then add freshly prepared BS2G to minimize local hydrolysis.

Comparison (literature/general):

• BS2G vs DSG (non‑sulfonated): BS2G is water‑soluble/membrane‑impermeant; DSG often requires organic cosolvent and can permeate membranes.
• BS2G vs BS3 (longer spacer): BS3 provides longer reach; BS2G affords more compact crosslinks with potentially higher structural restraint.
• BS2G vs EDC/NHS: BS2G is a pre‑activated, homobifunctional linker targeting amines; EDC/NHS creates zero‑length amide bonds between carboxylates and amines and is highly buffer‑dependent.

Storage and Reconstitution

Item-specific conditions (from Product Data):

• Storage: Store at −20°C.
• Shipping: Shipped in an ice chest with ice pads.
• Research use note: For research use only.

General/literature best practices for sulfo‑NHS crosslinkers:

• Keep tightly sealed under dry, inert atmosphere (desiccant recommended). Minimize headspace humidity; moisture triggers hydrolysis and loss of activity.
• Aliquot on receipt into small, single‑use vials under dry conditions to avoid repeated warming and moisture ingress.
• Protect from light if supplied in amber or opaque packaging.

Reconstitution (literature/general):

• Prepare solutions immediately before use. Dissolve in high‑purity water or compatible buffer (pH 7.2–8.5). Alternatively, prepare a concentrated stock in anhydrous DMSO or DMF and add to the reaction last to achieve final working concentration.
• Keep solutions on ice and use within a short timeframe (typically minutes to an hour depending on pH/temperature). Discard remaining solution after use; do not refreeze aqueous solutions.
• Avoid amine‑containing buffers during dissolution and reaction. After coupling, quench with Tris or glycine as needed.

Any item‑specific limits for water content, stabilizers, or shelf life: Not specified for this item; refer to CoA/Spec Sheet and SDS for definitive guidance.

Structure and Identity

Item-specific identifiers provided in Product Data are limited; key structural details are summarized carefully below.

• Product name: BS2G Crosslinker – disodium (homobifunctional NHS-ester crosslinker)
• CAS: 910292-86-3 (from Product Data)
• 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: Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey: Not specified for this item; refer to CoA/Spec Sheet.

Structural features (general/literature description):

• BS2G is widely known as the disodium salt of bis(sulfosuccinimidyl) glutarate, i.e., a homobifunctional N‑hydroxysulfosuccinimide (sulfo‑NHS) diester of glutaric acid. Each terminus contains an activated sulfo‑NHS ester that selectively reacts with nucleophilic primary amines (e.g., ε‑NH2 of lysine) to form stable amide bonds (literature/general).
• The central spacer corresponds to a glutarate (five‑carbon) diacid backbone, giving a relatively short, well‑defined inter-amine distance upon crosslinking (literature/general).
• As a disodium salt of the sulfo‑NHS form, BS2G is typically water‑soluble and membrane‑impermeant, favoring surface and extracellular bioconjugations (literature/general).

2D structure in words (literature/general):

• Symmetric diester: sulfo‑NHS carbonyl‑activating group – carbonyl – (CH2)3 – carbonyl – sulfo‑NHS.
• Ionic sulfonate substituents (neutralized by Na+) on each NHS ring confer hydrophilicity. No stereogenic centers; overall molecule is achiral but amphiphilic due to ionic sulfonates balanced by hydrophobic succinimide rings.

Synthetic Utility

From a synthetic chemistry standpoint, BS2G is a bifunctional amine‑reactive acylating agent that delivers a defined glutarate spacer between two nucleophiles.

Utility (literature/general):

• Homobifunctional linker: Facilitates formation of inter‑ or intramolecular diamide bridges between primary amines on small molecules, peptides, and polymers.
• Orthogonal strategies: Can be combined with thiol‑selective chemistries (maleimide, haloacetyl), click reactions (azide–alkyne), or enzymatic ligation to build complex conjugates in a stepwise manner where BS2G introduces amide‑based tethers selectively at amines.
• Materials chemistry: Used to crosslink amine‑rich coatings (chitosan, poly(allylamine), PEI) or to couple amine‑functional nanoparticles to proteins, enhancing stability and controlling spacing.

Mechanistic notes (literature/general):

• The sulfo‑NHS leaving group activates the carboxylate toward aminolysis; the rate depends on amine pKa, sterics, and microenvironment. Hydrolysis competes with aminolysis; thus, kinetics often follow pseudo‑first‑order behavior under excess amine.
• The glutarate spacer imparts moderate flexibility and a short reach, which can enhance structural restraint and aid interpretation in crosslinking mass spectrometry (XL‑MS) compared to longer linkers.

Item‑specific reactivity data or kinetic constants: Not specified for this item; refer to CoA/Spec Sheet and primary literature.

Target Specificity

Not an antibody or biological targeting reagent with predefined antigen/epitope specificity.

• Mechanistic selectivity (literature/general): Reacts broadly with primary aliphatic amines (e.g., lysine ε‑amines, N‑termini) under mildly basic conditions. Secondary amines react more slowly; thiols and alcohols are generally unreactive under standard conditions. Thus, BS2G exhibits chemical, not biological, specificity.

Item‑specific antigen/epitope data: Not applicable.

Häufig gestellte Fragen

How should this product be stored?

Store at ?20 °C. Freezer storage is required to maintain the specified shelf life.

How is this product shipped?

This product ships in an insulated container with ice pads. Unpack on arrival and transfer it to the storage condition stated above.

What are the CAS number, molecular formula and molecular weight?

The CAS Number is 910292-86-3, the molecular formula is C13H10D4N2O8, and the molecular weight is 330.28 g/mol.

What documentation is provided?

Available product documentation, including Certificates of Analysis (COA), Safety Data Sheets (SDS), and specification sheets, is shown in the product document area. Document availability and access follow the current site policy.