Tris Antigen Retrieval Solution (10×, pH 9.0) - BioReagent,for microscopy,Suitable for Immunohistochemistry(IHC),10×

Cat. No.: T1508976

GRADE & PURITY BioReagent, for Microscopy, Suitable for Immunohistochemistry(IHC), 10×

Synonyms

Tris Antigen Retrieval Solution

Storage

Room temperature

Shipped In

Normal

Application

Cell Staining, IHC

Vue d’ensemble

  During tissue preparation, antigens are masked by the effects of chemical reagents, and the peptide chains of some antigens become distorted due to heating. As a result, these antigens cannot be visualized during immunohistochemical staining. To address these issues, the process of re-exposing or restoring these antigens using chemical reagents and heat is called antigen retrieval. Buffers such as citrate, EDTA, or Tris can re-expose antigens masked by formalin under heating conditions without damaging antigenic epitopes, thereby improving antigen detection rates, reducing background staining, and enhancing diagnostic accuracy.

  Tris Antigen Retrieval Solution (10×, pH 9.0) is composed of Tris, a trace amount of Tween 20, and other ingredients. It can effectively eliminate protein cross-linking caused by aldehyde-based fixatives and fully expose antigenic epitopes in samples such as paraffin sections and frozen sections. This solution is suitable for antigen retrieval of samples (including paraffin sections and frozen sections) fixed with paraformaldehyde, formaldehyde, or other aldehyde-based reagents. Antigen retrieval can improve the immunostaining effect of paraffin sections and enhance the staining effect of frozen sections to varying degrees. When the immunostaining effect of frozen sections is unsatisfactory, antigen retrieval is recommended. Based on a dosage of 10 ml of Tris Antigen Retrieval Solution (1×) per slide, 100 ml of the 10× concentrated solution can be used for antigen retrieval of 100 samples. This reagent is intended for research purposes only and not for clinical diagnosis or other applications.

Materials to Be Prepared by Users

1. Graded ethanol solutions

2. Double-distilled water or deionized water

3. Heating equipment

4. Immunostaining wash buffer

Operating Procedures (For Reference Only)

(1) Paraffin Sections

1. Deparaffinization and Rehydration

① Deparaffinize sections with xylene or deparaffinization clearing solution for 3 times, 3-5 minutes each time.

② Dehydrate with anhydrous ethanol for 2 times, 3-5 minutes each time.

③ Treat with 95% ethanol for 3-5 minutes.

④ Treat with 90% ethanol for 3-5 minutes.

⑤ Treat with 80% ethanol for 3-5 minutes.

⑥ Treat with 70% ethanol for 3-5 minutes.

⑦ Rinse with distilled water for 2 times, 3-5 minutes each time.

2. Antigen Retrieval

① Dilute Tris Antigen Retrieval Solution (10×, pH 9.0) to 1× working concentration with deionized water or double-distilled water.

② Immerse sections in the 1× Tris Antigen Retrieval Solution, and heat at 95 °C or boiling temperature for approximately 10-30 minutes.

③ Preheat the 1× Tris Antigen Retrieval Solution to 95-100 °C before use. If heating with a microwave oven, avoid boiling and excessive water evaporation. Then allow the solution to cool down to room temperature naturally within approximately 20-30 minutes.

3. Wash sections with immunostaining wash buffer for 1-2 times, 3-5 minutes each time.

4. Proceed with subsequent immunostaining steps, such as blocking.

(2) Frozen Sections

1. Dilute Tris Antigen Retrieval Solution (10×, pH 9.0) to 1× working concentration with deionized water or double-distilled water.

2. Wash sections with immunostaining wash buffer for 5 minutes.

3. Immerse sections in the 1× Tris Antigen Retrieval Solution, and heat at 95 °C or boiling temperature for approximately 10-30 minutes.

4. Preheat the 1× Tris Antigen Retrieval Solution to 95-100 °C before use. If heating with a microwave oven, avoid boiling and excessive water evaporation. Then allow the solution to cool down to room temperature naturally within approximately 20-30 minutes.

5. Wash sections with immunostaining wash buffer for 1-2 times, 3-5 minutes each time.

6. Proceed with subsequent immunostaining steps, such as blocking.

(3) Other Samples

Follow the procedures for paraffin sections or frozen sections.

Precautions

1. The optimal heating time for sections immersed in the 1× Tris Antigen Retrieval Solution should be determined empirically according to different samples and target proteins.

2. For your safety and health, wear a lab coat and disposable gloves during operation.

3. Use the reagent as soon as possible after opening to avoid compromising subsequent experimental results.

Specifications

Synonymes

Tris Antigen Retrieval Solution

Spécifications et pureté

BioReagent,for microscopy,Suitable for Immunohistochemistry(IHC),10×

Conditions de stockage de stockage

Room temperature

Expédié en

Normal

Grade

BioReagent, pour la microscopie, Suitable for Immunohistochemistry(IHC)

Product Line

Produits biologiques

Noms et identifiants

PH

9.0

Application Protocols

General protocol for Heat-Induced Epitope Retrieval (HIER) using Tris Antigen Retrieval Solution (10×, pH 9.0). Adjust parameters per antibody vendor recommendations.

• Preparation

  • Dilute the 10× stock 1:10 with deionized water to obtain 1× working solution. Mix gently; verify clarity and pH if required.

• Deparaffinization and rehydration

  • Bake slides (e.g., 60 °C, 30–60 min). Deparaffinize in xylene (2×, 5–10 min each), then rehydrate through graded ethanols to water.

• Retrieval

  • Pre-warm the 1× buffer. Immerse slides fully in a heat-safe vessel.
  • Heat to 95–100 °C for 10–30 minutes (optimize per antibody/tissue). Maintain consistent temperature throughout.
  • Allow slides to cool in the buffer for 10–20 minutes to near room temperature.

• Rinsing and IHC workflow

  • Rinse slides in TBS or PBS (3×, 5 min). Proceed to block endogenous peroxidase (if HRP/DAB), then protein blocking, primary antibody incubation, detection, and counterstaining.

• Tips and QC

  • Use charged slides to minimize tissue detachment at high pH/temperature.
  • Ensure adequate buffer volume (≥300–500 µL per slide in small jars; proportionally more in dishes) and avoid boil-dry.
  • Include tissue controls and record instrument temperature profiles.

Note: This is general guidance. It does not supersede antibody-specific instructions or your laboratory’s validated SOPs.

Biological Roles

This section addresses the buffer’s role in biochemical context; it is not a therapeutic description.

• Tris buffer in biochemistry (general/literature)

  • Tris (tris(hydroxymethyl)aminomethane) is a widely used biological buffer with a pKa near 8.06 at 25 °C, providing effective buffering between ~7.5 and 9.0.
  • The buffering mechanism derives from protonation/deprotonation of its primary amine; the pKa is temperature dependent, which is critical in heated applications like antigen retrieval.
  • In histology, alkaline buffers modulate protein charge states and hydrogen-bonding networks, promoting partial reversal of formalin-induced crosslinks and improving epitope exposure.

• Compatibility considerations (general)

  • Tris can interact with certain enzymes, metal ions, and aldehyde residues; for antigen retrieval, such interactions are typically acceptable or beneficial.
  • Tris buffers can interfere with some downstream labeling chemistries (e.g., certain amine-reactive crosslinkers), but standard IHC detection systems (HRP/DAB, AP/Vector Red, polymer-based) are generally compatible after retrieval and appropriate washing.

Buffer Applications

Designed specifically for antigen retrieval in IHC; broader buffer system uses are noted for context.

• Intended use (item-specific)

  • 10× Tris Antigen Retrieval Solution at pH 9.0 for HIER on FFPE tissue sections; dilute 1:10 with deionized water to make 1× working solution.

• Operating window (general/literature)

  • Tris buffering is optimal near pH 7.5–9.0 (pKa ~8.06 at 25 °C). The alkaline setting (pH 9.0) is suited for antigens favoring basic retrieval.
  • Temperature strongly influences effective pH; validate conditions at actual retrieval temperature (often 95–100 °C in HIER instruments).

• Practical recipes (general guidance; not item specifications)

  • 1× working solution: Prepare by diluting the 10× stock 1:10 with high-purity water. Mix gently and verify clarity.
  • Optional additives: Some labs add nonionic detergents (e.g., 0.05% Tween 20) to enhance wetting; these are not part of this item unless specified in the CoA and should be validated.

• Use tips for IHC (general best practice)

  • Ensure full coverage of tissue; avoid buffer boiling dry.
  • Post-retrieval: Cool slides in the buffer, then rinse in TBS or PBS before proceeding to blocking and primary antibody incubation.
  • Include positive and negative control tissues to confirm retrieval performance and antibody specificity.

Green Alternatives

The solution is already an aqueous, nonflammable buffer, aligning well with green chemistry principles. Considerations and alternatives are about optimizing performance and resource use rather than solvent substitution.

• Environmental & safety profile (general)

  • Water-based, minimal VOC emissions, nonflammable. Typically low acute hazard.

• Alternatives and trade-offs (general/literature)

  • Citrate buffer (pH ~6.0): Lower pH, effective for many targets; may require higher temperatures or longer times for difficult epitopes.
  • Tris-EDTA (pH ~8–9): Increased retrieval potency via chelation; may risk tissue lifting or morphology changes in delicate samples.
  • Proprietary high-pH buffers (pH 8.5–9.5): Similar performance window; composition may include detergents that increase waste treatment complexity.

• Resource efficiency tips (applicative)

  • Prepare only the needed volume of 1× working solution from the 10× stock to minimize waste.
  • Reuse 1× retrieval buffer across a limited number of runs only if validated in-house and contamination is controlled; performance can decline due to pH drift and extracted tissue components.
  • Employ closed heating systems (pressure retrievers) to reduce evaporation and energy consumption compared to open boiling.

Pharmaceutical Uses

Not typically applicable. This product is a laboratory-formulated antigen retrieval buffer for histology/IHC workflows and is labeled For research use only. It is not intended for manufacturing of drug products, clinical diagnostics, or therapeutic applications.

• General/literature context (non-item-specific)
  • Tris buffers are widely used in bioprocessing and formulations as pH control agents; however, suitability for GMP or pharmacopeial compliance requires product-specific documentation and qualification, which are not specified for this item.

For any regulated use, consult your quality unit and request detailed specifications and compliance documentation; otherwise, restrict use to research and microscopy.

Physical Properties

• Item-specific physical data

  • Concentration: 10× stock (dilute 1:10 to prepare 1× working solution)
  • pH: 9.0 (at 10×, as labeled)
  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Density, refractive index, conductivity, osmolality: Not specified for this item; refer to CoA/Spec Sheet.

• General/literature expectations for Tris buffers (context only; not item specifications)

  • Physical state: Aqueous solution; viscosity and density close to water, modestly concentration-dependent.
  • Buffering range: Effective around pH 7.5–9.0 (Tris pKa ~8.06 at 25 °C; temperature sensitive, ΔpKa/ΔT ≈ −0.028 per °C, literature).
  • Solubility: Fully miscible with water; not miscible with nonpolar solvents.
  • Temperature effects: Alkalinity increases with cooling (apparent pH rises as temperature decreases due to Tris pKa shift).

• Practical implications (general)

  • Calibrate pH at the intended working temperature for most reproducible antigen retrieval.
  • Use freshly prepared 1× working solution for consistent results; avoid CO2 absorption over long open exposure, which can slightly acidify unsealed alkaline buffers.

Quality and Grades

• Item-specific grade attributes

  • Grade: BioReagent, for Microscopy; Suitable for Immunohistochemistry (IHC)
  • Implication: Formulated for consistent staining performance, low background, and compatibility with common HIER workflows on FFPE sections.

• What the grade means (general guidance)

  • BioReagent: Manufactured with raw materials and processes suitable for biological research, emphasizing low bioburden and consistent formulation.
  • For Microscopy: Meets expectations for optical clarity (no precipitates/particulates) to minimize slide artifacts and supports reproducible morphology preservation during HIER.
  • Suitable for IHC: Validated to be compatible with antibody-based detection workflows, minimizing epitope damage and background developed under alkaline retrieval conditions.

• Stabilizers/preservatives

  • Not specified for this item; refer to CoA/Spec Sheet for any antimicrobial preservative or chelator content.

• Quality controls (general best practices)

  • Lot-to-lot verification of pH (at stated temperature) and concentration.
  • Visual inspection for clarity prior to use; discard if turbidity or precipitate forms, which can indicate contamination or evaporation.
  • Performance verification can include running a known positive control tissue and confirming expected staining intensity after standardized HIER.

Reaction and Applications

Primary application: Heat-Induced Epitope Retrieval (HIER) for immunohistochemistry on FFPE tissues.

• Mechanistic context (general/literature)

  • Formalin fixation creates methylene bridges and crosslinks that mask protein epitopes. Alkaline Tris buffer at elevated temperature facilitates hydrolysis/reversal of some crosslinks and disrupts noncovalent interactions, restoring antibody accessibility.
  • pH 9.0 favors deprotonation of side chains (e.g., Lys, Tyr) and modulates protein conformation, aiding unmasking of select epitopes.

• Practical advantages (application-focused)

  • Broad compatibility with primary antibodies that specify “alkaline retrieval” conditions.
  • Often improves nuclear antigen staining (e.g., transcription factors) and some membrane markers when citrate pH 6.0 is suboptimal.
  • Balanced retrieval strength versus EDTA buffers: less chelation-driven extraction of metal ions, which can preserve morphology in certain tissues.

• Implementation notes (general best practice)

  • Use positively charged or adhesion slides to minimize tissue lifting at high temperature and basic pH.
  • Ensure even heating (pressure cooker, decloaking chamber, microwave with temperature control, or water bath). Target solution temperature typically 95–100 °C for HIER.
  • Pre-warm buffer and maintain sufficient volume (≥300–500 µL per slide in coplin jars; more in staining dishes) to ensure complete coverage.
  • After retrieval, allow a controlled cool-down (10–20 min) in buffer to reduce sudden conformational changes and tissue detachment.

Note: This product is not intended for synthetic chemical reactions; its value lies in histological sample preparation.

Reaction Conditions

Framed for HIER process conditions rather than chemical synthesis.

• Typical HIER conditions with Tris pH 9.0 (general/literature guidance)

  • Working solution: 1× (prepared by 1:10 dilution of the 10× stock with deionized water).
  • Temperature: 95–100 °C in a water bath, steamer, or dedicated pressure retrieval system; some protocols use sub-boiling 90–95 °C for sensitive tissues.
  • Time: 10–30 minutes at target temperature, followed by 10–20 minutes cool-down in buffer. Optimize per antibody datasheet.
  • Vessels: Heat-resistant staining dishes, Coplin jars, or pressurized decloaking chambers; ensure adequate buffer volume to fully immerse slides.
  • Post-treatment: Rinse in TBS or PBS to normalize ionic environment before blocking and antibody incubation.

• Factors influencing outcomes (general)

  • Tissue fixation severity (time in formalin, crosslink density) and section thickness.
  • Slide adhesion quality; high-pH/high-temperature conditions increase risk of tissue detachment—use charged slides and proper baking.
  • pH/temperature calibration: Tris pKa is temperature-dependent; confirm performance at actual operating temperature.

Note: Times/temperatures are general literature guidance for HIER and not product-specific specifications. Validate conditions for each antibody–tissue pairing.

Safety and Handling

• Item-specific hazard data

  • GHS classification, signal word, pictograms, H-statements: Not specified for this item; refer to the SDS.

• General safety guidance (for Tris-based aqueous buffers; literature/general)

  • Expected hazards: Typically low hazard; may cause mild eye/skin irritation. Avoid ingestion and inhalation of aerosols/mists.
  • PPE: Lab coat, safety glasses, and nitrile gloves. Use splash protection when heating during HIER.
  • Incompatibilities: Strong acids (neutralization/heat), strong oxidizers. Avoid contact with reactive acid chlorides/anhydrides.
  • Thermal use: Antigen retrieval involves near-boiling temperatures or pressure vessels; use heat-resistant gloves, secondary containment, and ensure vessels are rated for temperature and pressure.
  • First aid overview: In case of eye/skin contact, rinse with water for several minutes; remove contaminated clothing. If inhaled (mists), move to fresh air. If ingested, rinse mouth and seek medical advice. Always follow institutional SOPs and SDS.
  • Spill response: Absorb with inert material, then water-rinse the area. For large volumes, contain and collect as aqueous nonhazardous waste per local regulations unless experimental contaminants are present.

Always consult the product-specific SDS for authoritative hazard, exposure limits, and disposal requirements.

Solvent Selection

This product is an aqueous buffer; solvent selection is not typically variable within its intended use. Key considerations:

• Polarity and miscibility (general)

  • Fully water-based and fully miscible with water; not miscible with nonpolar organic solvents.
  • Designed to be used as-is (after dilution to 1×) without co-solvents.

• When to choose this buffer (application-focused)

  • Select alkaline Tris pH 9.0 retrieval when targeting epitopes known to unmask better under basic HIER (e.g., many nuclear antigens and some membrane proteins), compared with citrate pH 6.0.
  • Prefer Tris pH 9.0 when EDTA-based high-pH buffers are too aggressive for delicate tissues yet citrate pH 6.0 is insufficient.

• Comparison to alternatives (general guidance)

  • Citrate buffer (pH ~6.0): Gentler, effective for many antigens but may under-retrieve some targets.
  • Tris-EDTA (pH ~9.0): Stronger chelation can enhance retrieval for calcified or crosslinked targets but may increase tissue detachment if slides are not positively charged.
  • Proprietary high pH retrieval solutions (pH 8.5–9.5): Similar application space; confirm compatibility with detection chemistry.

Storage and Reconstitution

• Item-specific storage

  • Storage conditions: Room temperature (per Product Data). Avoid extreme temperatures and protect from contamination. Shipped under normal conditions.

• Reconstitution/preparation

  • The product is supplied as a 10× concentrate. Prepare 1× working solution by diluting 1:10 with deionized or distilled water. Mix gently to avoid foaming.
  • If particulate matter is observed, do not use; clarify by filtration only if validated and sterile technique is required.

• Stability and handling (general guidance; not item-specific specs)

  • Many Tris buffers are stable for months at room temperature when unopened. After opening, minimize air exposure and re-cap promptly to limit CO2 absorption and evaporation.
  • Prepare fresh 1× solution as needed; if storing 1×, keep in a clean, closed container. For best consistency, use within 1–2 weeks or per in-house validation.
  • Do not freeze unless specified by the CoA/Spec Sheet.

• Research use

  • For research use only. Inspect solution clarity prior to each use and document lot numbers and preparation dates in your IHC records.

Structure and Identity

Concise description: This product is an aqueous, ready-to-dilute 10× Tris-based antigen retrieval buffer adjusted to pH 9.0 for heat-induced epitope retrieval (HIER) in immunohistochemistry.

• Item-specific facts (from Product Data)

  • Product: Tris Antigen Retrieval Solution (10×, pH 9.0)
  • Format: Concentrated buffer (10×)
  • Intended use: Immunohistochemistry (IHC) antigen retrieval
  • Storage: Room temperature; shipped under normal conditions
  • Grade: BioReagent, for Microscopy; Suitable for IHC
  • CAS/SMILES/InChIKey/Molecular formula/MW: Not specified for this item; refer to CoA/Spec Sheet.

• Structural features (general/literature for context; not item-specific specs)

  • Principal buffering component: Tris (tris(hydroxymethyl)aminomethane), a primary amine bearing three hydroxymethyl groups on a central carbon.
  • Functional groups: One primary amine (–NH2) and three alcohols (–CH2OH), providing a zwitterion-capable, hydrogen-bonding scaffold.
  • Buffering chemistry: Protonation/deprotonation of the primary amine controls pH in the alkaline near-neutral range.

• 2D structure in words (general/literature)

  • A central carbon attached to three –CH2OH substituents and one –NH2 substituent; typically supplied as free base or protonated salt in buffers.

Note: This is a formulated aqueous buffer system; the precise composition (e.g., Tris concentration, presence of chelators/surfactants, preservatives) is not specified for this item and should be confirmed by the CoA/Spec Sheet.

Synthetic Utility

Not typically applicable. This product is an aqueous Tris-based buffer formulated for antigen retrieval in IHC and is not intended as a reagent for organic synthesis or preparative chemical transformations.

• General note (literature)
  • Tris (the base) can act as a proton buffer and occasionally as a nucleophilic amine in chemical systems, but the formulated aqueous buffer is optimized for histological sample preparation, not for synthetic reactions.

For chemical synthesis needs, select reagents and solvents specifically intended for stoichiometric or catalytic transformations.

Target Specificity

Not applicable to this product. This is a buffer solution for antigen retrieval and does not have antigen or epitope specificity, nor does it include antibodies or targeting moieties. Consult the primary antibody datasheet for target-specific details.

Foire aux questions

What does for microscopy mean?

for microscopy indicates that the material has been processed and tested for specialty industrial processing. Specifications focus on the impurities that affect performance in that application — for example, metal content for electronics, particle size for batteries, refractive index for optics, or moisture content for moisture-sensitive processes.

What does BioReagent mean?

BioReagent indicates that the product has been processed and tested for biological and biochemical applications. Specifications may include endotoxin limits, microbial controls, sterility, low DNase/RNase/protease activity, and verified biological activity where relevant.

How should this product be stored?

Store at room temperature.

How is this product shipped?

This product ships under standard ambient conditions. No temperature-controlled packaging is required.

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.