Tris Buffered Saline - pH 8.0, powder

Cat. No.: T475659

GRADE & PURITY pH 8.0, powder

Synonyms

TBS

Storage

Room temperature

Overview

Contents of one pouch, when dissolved in one liter of distilled or deionized water, will yield 0.05 M Tris buffered saline (NaCl - 0.138 M; KCl - 0.0027 M); pH 8.0, at 25 °C.

Specifications

Synonyms

TBS

Specifications & Purity

pH 8.0, powder

Storage

Room temperature

Names and Identifiers

PH

8.0

Application Protocols

Item-specific, validated protocols are Not specified for this item; refer to CoA/Spec Sheet. The following are literature-based examples intended as starting points.

1. Western blot wash and antibody dilution (TBST)

• Prepare TBST: 1× TBS + 0.1% (v/v) Tween-20.
• Blocking: 1 h at RT in TBST + 5% nonfat dry milk or 3% BSA.
• Primary antibody: Dilute in TBST + 1–5% blocker; incubate 1–16 h (RT or 4 °C).
• Wash: 3–5 × 5–10 min in TBST.
• Secondary antibody: Dilute similarly; incubate 1 h; wash as above; develop.

2. ELISA plate washing

• Wash buffer: TBS or TBST (0.05% Tween-20) pre-warmed to RT.
• Perform 3–5 wash cycles between steps; allow 30–60 s soak per cycle.

3. IHC/ICC rinses

• Rinse sections/cells 3 × 5 min with TBS between blocking, antibody, and detection steps; adjust pH near 7.4–7.6 per protocol.

4. Sample/column handling

• Equilibrate affinity or ion-exchange columns with TBS when phosphate interferes; match ionic strength to sample.

Note: For amine-reactive conjugations (EDC/NHS, succinimidyl esters), switch to amine-free buffer prior to activation/labeling, then return to TBS after quenching (literature guidance).

Biological Roles

TBS functions as a controlled physicochemical environment for biological macromolecules rather than exerting a biological effect per se.

Mechanistic roles (literature; general):

• pH control: Tris/Tris·HCl constitutes a conjugate acid–base pair with pKa ≈ 8.06 at 25 °C, providing effective buffering from ~pH 7–9. This stabilizes protein structure and antibody–antigen interactions during immunoassays and washes.
• Ionic strength: NaCl modulates electrostatic interactions, influencing protein solubility, adsorption to membranes/plates, and nonspecific binding. Adjustments in salt concentration alter assay stringency.
• Temperature effects: Tris pKa decreases with increasing temperature (~0.028–0.03 pH/°C), so apparent pH at room temperature vs cold-room vs 37 °C will differ unless corrected, impacting enzyme activity and binding equilibria.
• Metal binding: Tris is a weak chelator; typically minimal biological impact, but it can perturb certain metalloenzyme assays at high concentrations.

Use cases (general):

• As a wash/diluent in Western blotting, IHC/ICC, ELISA, and general protein handling when phosphate is undesirable (e.g., alkaline phosphatase detection systems).

No medical or clinical claims: This product is intended for research use only (as stated by the manufacturer). Item-specific composition and performance characteristics are Not specified for this item; refer to CoA/Spec Sheet.

Buffer Applications

Tris Buffered Saline is widely used as an assay and wash buffer. Exact formulation for SKU T475659 is Not specified for this item; refer to CoA/Spec Sheet. The following are literature-standard examples for planning purposes only.

Typical formulations (literature examples):

• 1× TBS, pH 7.6: 20 mM Tris, 150 mM NaCl; adjust pH with HCl at 25 °C.
• 10× TBS stock: 200 mM Tris, 1.5 M NaCl; dilute to 1× before use.
• TBST (for washing): 1× TBS + 0.05–0.1% (v/v) Tween-20.

Practical guidance:

• pH calibration: Adjust at the temperature of use. For cold-room operations, consider setting pH slightly higher at room temperature to achieve the target at 4 °C (accounting for Tris’s temperature coefficient).
• Stringency control: Increase NaCl to reduce nonspecific binding in immunoassays; reduce salt for weaker interactions.
• Sterility: For sterile applications, 0.2 μm filter-sterilize and store in clean, closed containers. Consider preservative-free formulations for peroxidase-based assays and live cells.
• Compatibility: Avoid Tris-containing buffers for amine-reactive conjugation chemistries (EDC/NHS, succinimidyl esters).

Recipes and QC parameters (conductivity, osmolality, endotoxin) for this specific product are Not specified for this item; consult the product label/CoA.

Green Alternatives

TBS is an aqueous buffer and generally aligns with green chemistry principles for benign media. Environmental impact considerations derive from production purity, packaging, and any added preservatives.

Greener-choice discussion (general):

• Aqueous base: Water as solvent is inherently preferable to organic solvents for washing/dilution steps.
• Preservatives: Sodium azide and certain isothiazolinones are hazardous to aquatic life. Preservative-free, single-use sterile formats reduce biocide use but may increase packaging waste—tradeoff to consider.
• Concentrates/tablets: Shipping concentrates (e.g., 10×) or tablets reduces transport mass/volume and packaging, improving sustainability relative to shipping large volumes of 1×.

Comparison (literature; general):

• TBS vs PBS: Similar environmental profiles; PBS avoids amine-containing components but introduces phosphate, which can contribute to eutrophication if discharged untreated.
• TBS vs HEPES-buffered saline: HEPES is more complex to synthesize; choose only when performance benefits (stable pH with temperature, cell culture) justify the footprint.

Best practices:

• Prepare only the amount needed; store properly to minimize microbial spoilage and waste.
• Dispose according to local regulations—especially if the formulation contains preservatives (check CoA/SDS).

Item-specific composition and preservative content: Not specified for this item; refer to CoA/Spec Sheet.

Pharmaceutical Uses

This product is supplied for research use only. It is not intended for human or veterinary use, diagnostic procedures, or clinical applications.

Industry context (general):

• Process buffers: Tris-buffered saline is commonly employed during upstream/downstream bioprocessing steps (e.g., column equilibration/wash, intermediate hold solutions) where phosphate is undesirable. In GMP settings, composition, bioburden, and endotoxin limits are tightly specified and validated.
• Formulation excipient: While Tris and saline components appear in some authorized drug product formulations, any use in finished dosage forms requires pharmacopeial-grade materials and process controls beyond research-grade offerings.
• Device/assay manufacturing: TBS may be used in IVD reagent manufacturing as wash/diluent; preservative content and surfactant levels are tailored to assay design.

For SKU T475659, grade, sterility, endotoxin, and preservative content are Not specified for this item; refer to CoA/Spec Sheet. Users operating under regulated frameworks should verify compliance with relevant pharmacopeial and quality standards before use.

Physical Properties

This product is a buffered saline mixture; item-specific physico-chemical specifications (pH, concentration, conductivity, osmolality, etc.) are Not specified for this item; refer to CoA/Spec Sheet.

Typical properties of Tris-buffered saline (literature; for context only):

• Physical state: Aqueous solution (or concentrate/tablets in some formats); clear, colorless.
• Typical pH ranges: ~7.2–8.0 when formulated with Tris/Tris·HCl; common setpoints 7.4 or 7.6 (25 °C). Strong temperature dependence: Tris pKa decreases ~0.028–0.03 pH units/°C above 25 °C.
• Buffering range (Tris, literature): pKa ≈ 8.06 at 25 °C; effective range ~pH 7–9.
• Ionic strength/osmolality: depends on NaCl concentration (commonly 100–150 mM NaCl at 1×; osmolality roughly 200–300 mOsm/kg at 1×; literature values).
• Density, refractive index, viscosity: close to water at the same temperature for 1× solutions (literature).
• Solubility: Miscible with water; not miscible with nonpolar solvents.

Practical implications (general):

• pH drifts with temperature; adjust and report at the working temperature.
• Elevated ionic strength enhances stringency in immunoassays and can reduce nonspecific binding; conversely, lower salt may increase background interactions.

All numeric values above are literature guidance only and not specifications for SKU T475659.

Quality & Grades

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

Context for buffer grades (general guidance):

• Molecular biology grade: Prepared with high-purity reagents and water (18 MΩ·cm), typically DNase/RNase-free; suitable for nucleic acid/protein workflows where nuclease contamination is critical.
• Cell culture grade: Sterile-filtered (e.g., 0.2 μm), low endotoxin; may be gamma-irradiated or aseptically filled. Specifications would include bioburden and endotoxin limits.
• Electrophoresis/Western blot grade: Focus on low background fluorescence/absorbance; consistent ionic strength for reproducible transfer and washing.
• Preservatives: Some TBS products include sodium azide or other biocides to limit microbial growth; others are preservative-free for compatibility with peroxidase-based detection or live-cell contexts. Presence/absence is not specified for SKU T475659.

What to check on the CoA/Spec Sheet:

• Format (ready-to-use 1× vs. concentrates/tablets), target pH and tolerance, conductivity/osmolality, sterility status, preservative content, and permissible microbial/endotoxin limits.

Selection note:

• Align grade with intended downstream application (e.g., AP-based detection favors TBS; peroxidase compatibility may require azide-free formulations).

Reaction & Applications

Primary use domains (general, life-science workflows):

• Immunoblotting (Western blot): As wash buffer and antibody diluent; TBST (TBS + Tween-20) reduces nonspecific binding.
• Immunohistochemistry (IHC) and immunocytochemistry (ICC): For tissue/cell washing and antibody dilutions at pH ~7.4–7.6 (literature). Tris can participate in certain epitope retrieval protocols when combined with other agents.
• ELISA: Plate washing and sample/antibody dilution; salt and detergent levels modulate assay stringency and background.
• Protein purification/handling: Maintaining protein solubility and activity during dialysis, column equilibration, and fraction collection when phosphate is undesirable.

Compatibility cautions (general):

• Amine reactivity: Free Tris base contains a primary amine, which can quench NHS-esters and interfere with carbodiimide (EDC/NHS) coupling and aldehyde-based conjugations (Schiff base). Use amine-free buffers (e.g., PBS, MES, HEPES) for bioconjugation steps.
• Metal interactions: Tris is a weak ligand; avoid if metal-catalyzed reactions or precise metal speciation are critical.
• Enzymes: Tris pH drift with temperature can influence enzyme kinetics; calibrate pH at the working temperature.

Practical tips (literature):

• Degas only if required for sensitive assays; routine degassing is usually unnecessary.
• For TBST, prepare fresh detergent-containing solutions or verify stability to prevent micelle-related variability.

Item-specific instructions and validated applications: Not specified for this item; refer to CoA/Spec Sheet.

Reaction Conditions

Because TBS serves primarily as an assay/wash medium, “reaction conditions” are best framed as operational parameters for common lab workflows (literature guidance; not item-specific):

• Western blot washing (TBST): 3 × 5–10 min washes at RT with gentle agitation in TBS + 0.1% Tween-20 after antibody incubations. Increase to 5 × 10 min for high-background membranes.
• Antibody dilution: Prepare primary/secondary antibodies in TBS or TBST with appropriate blockers (e.g., 1–5% BSA or milk, assay-dependent). Avoid Tris-based buffers for HRP substrates that are azide-sensitive if the TBS contains azide.
• ELISA plate washing: 3–5 cycles, 200–350 μL/well of TBS or TBST, with brief soak (30–60 s) per wash.
• IHC/ICC rinsing: 3 × 5 min TBS washes between steps; pH ~7.4–7.6 commonly used (literature), adjust to tissue/protocol requirements.
• Protein handling: Equilibrate columns and dilute samples with TBS when phosphate is contraindicated; match salt concentration to downstream steps to prevent precipitation or conformational changes.

Environmental/physical parameters:

• Temperature: Most washes at RT; cold-room (4 °C) for labile proteins. Remember Tris pH shift with temperature.
• Additives: Detergents (Tween-20), blockers (BSA, casein), and preservatives are added as needed; confirm compatibility with detection chemistry.

For SKU T475659, validated conditions and exact composition are Not specified for this item; consult the CoA/Spec Sheet.

Safety & Handling

GHS/SDS status for this specific item is Not specified for this item; refer to CoA/Spec Sheet and the product SDS for authoritative information.

General laboratory safety guidance for Tris-buffered saline (mixture):

• Expected hazards: Typically low hazard; however, composition (e.g., presence/absence of preservatives such as sodium azide or ProClin) is not specified and may alter hazard classification. Treat unknown formulations with caution.
• PPE: Wear lab coat, safety glasses, and appropriate chemical-resistant gloves. Avoid contact with eyes/skin and do not ingest.
• Handling: Use clean technique to avoid microbial contamination. If sterility is required, work aseptically and keep containers tightly closed. Avoid heating sealed containers.
• Incompatibilities (general):
  • Strong acids/bases can alter pH and buffer capacity.
  • Reactive acylating agents and NHS-esters can be quenched by Tris’s primary amine if present free in solution (see Synthetic Utility). Confirm formulation before use in conjugation workflows.
• First aid (general): Rinse skin/eyes with water for several minutes; remove contaminated clothing. If inhaled (aerosols), move to fresh air. If ingested, rinse mouth. Seek medical attention if irritation persists.
• Spills: Absorb with inert material, then water-rinse. Dispose according to institutional and local regulations.

Signal word, hazard statements, pictograms, and exact classification: Not specified for this item; consult the SDS.

Solvent Selection

Not a solvent to be selected for dissolving analytes in the organic synthesis sense; Tris Buffered Saline is itself an aqueous buffer medium.

When to choose TBS (general, literature-based):

• Immunoassays and blotting: Preferred when using alkaline phosphatase (AP)-conjugated antibodies because phosphate (in PBS) can inhibit AP. TBS maintains physiological ionic strength without phosphate.
• Protein handling: Suitable for protein dilution, washing, and storage when a slightly alkaline pH and defined ionic strength are needed.
• Surface/stringency control: Adjust NaCl concentration to tune ionic strength and reduce nonspecific interactions.

Comparison to common alternatives (general):

• TBS vs PBS: TBS avoids phosphate interference in AP-based detection; PBS provides phosphate buffering that can chelate or interact with some metal-dependent systems. pH temperature sensitivity is higher for Tris than for phosphate.
• TBS vs HEPES-buffered saline: HEPES has minimal temperature-dependent pKa drift and is often favored for live-cell work; TBS is economical and widely used for fixed-cell/tissue and protein applications.
• TBST: Addition of Tween-20 (e.g., 0.05–0.1% v/v, literature) increases washing stringency in Westerns and ELISAs.

Item-specific composition and pH: Not specified for this item; refer to CoA/Spec Sheet.

Storage & Reconstitution

• Storage conditions (from Product Data): Room temperature.
• Shipped in: Not specified for this item.
• Format (solution, concentrate, tablets) and reconstitution/dilution instructions: Not specified for this item; refer to CoA/Spec Sheet and product label.

General best practices (literature; for guidance only):

• Ready-to-use solutions: Store tightly closed at RT away from direct sunlight. If sterility is required, aliquot under aseptic conditions. Monitor for turbidity or microbial growth; discard if contamination is observed.
• Concentrates (e.g., 10×): Bring to RT, mix thoroughly before aliquoting; dilute with high-purity water (≥18 MΩ·cm). Verify pH after dilution and adjust at the working temperature.
• Tablets/powders: Dissolve each unit in the manufacturer-specified volume of water; mix until fully dissolved; adjust pH if instructed by the specification.
• Preservatives: Presence/absence is not specified. If preservative-free, consider cold storage for long-term stability or sterile filtration to extend shelf life.
• Freeze–thaw: Generally unnecessary for aqueous buffers; avoid freezing unless specified, as salt crystallization can shift composition upon thaw.

Research Use Only: As stated by the manufacturer, this product is for research use only.

Structure & Identity

Tris Buffered Saline (TBS) is a formulated aqueous buffer system, not a single defined molecule. It typically contains Tris (tris(hydroxymethyl)aminomethane) and sodium chloride, with pH adjusted using HCl (literature). Item-specific identifiers are not provided for this catalog entry.

• SKU: T475659
• Product Type: Pre-formulated buffer (mixture)
• CAS, CID, InChIKey, SMILES: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular Formula / Molecular Weight: Not applicable to the mixture. For reference (literature):
  • Tris base: C4H11NO3; MW 121.14 g/mol
  • NaCl: MW 58.44 g/mol
• Structural features (literature):
  • Tris contains a primary amine (–NH2) and three primary alcohols (–CH2OH), providing buffering capacity via the conjugate acid/base pair around physiological pH.
  • NaCl supplies ionic strength; no buffering functionality.
• 2D structural description (literature):
  • Tris: central quaternary carbon bearing one –NH2 and three –CH2OH substituents; no rings, no stereocenters.
  • Saline component is ionic, dissociated Na+ and Cl− in water.

Notes

• Exact composition, concentration (e.g., 1× vs 10×), and pH are Not specified for this item; refer to CoA/Spec Sheet.

Synthetic Utility

As a buffered saline medium, TBS is not typically a reagent in synthetic organic chemistry. However, its composition has important implications for bio-conjugation and biomaterials workflows.

Key considerations (general, literature):

• Amine interference: Tris contains a primary amine that competes with nucleophiles in amine-reactive chemistries (e.g., NHS-ester labeling, EDC/NHS coupling to carboxylates, aldehyde–amine (Schiff base) formation). Use amine-free buffers (PBS, MES, HEPES, carbonate/bicarbonate without primary amines) during coupling. TBS may be reintroduced after quenching.
• Enzyme-compatible milieu: Tris buffering near neutrality supports many enzyme-catalyzed transformations performed in aqueous media; verify pH/ionic strength requirements.
• Electrostatics management: Ionic strength (NaCl) helps control adsorption of biomolecules to supports (membranes, resins) during immobilization/cleavage steps.
• Metal-catalyzed systems: Tris can weakly coordinate metals; if precise metal speciation or catalytic turnover is crucial, consider non-coordinating buffers.

Workflow strategy:

• For conjugations: Exchange into amine-free buffer prior to activation/labeling (e.g., desalting column, dialysis), perform the reaction, then exchange back into TBS for storage/assay operations.

Item-specific composition (pH, salt) that would affect reactivity is Not specified for this item; refer to CoA/Spec Sheet.

Target Specificity

Not applicable. This product is a general-purpose buffer and is not an antibody, enzyme, or ligand with defined biological target specificity.

• Antigen/epitope, clone, isotype, and species reactivity: Not applicable.
• Any application-specific binding characteristics would derive from reagents used alongside TBS (e.g., antibodies), not from TBS itself.