Thyroxine-KLH (T4-KLH)

Cat. No.: T1522909

GRADE & PURITY BioReagent, Suitable for molecular biology

Panoramica

T4/KLH is an antigen-adjuvant conjugate formed by the coupling of T4 (L-Thyroxine) with keyhole limpet hemocyanin (KLH). By coupling the antigen with the protein adjuvant, the production of antigen-specific antibodies in vaccine models can be enhanced. The conjugate does not affect protein folding or disrupt major epitopes, and it can enhance cross-presentation and the generation of antigen-specific T cells.

Product Overview
Target	Thyroxine
Nature	Synthetic
Conjugate	mcKLH
Format	Liquid
Concentration	Please refer to the vial lablel for the specific concentration.
Buffer	0.01M pH7.4 PBS.
Storage	Aliquot and store at -20 °C. Avoid repeated Freeze/Thaw cycles. Store in the Dark.
Restrictions	For Research Use Only. Not for use in diagnostic procedures.

Specifications

Product Name

Thyroxine-KLH (T4-KLH)

Sinonimi

Thyroxine T4-KLH | Thyroxine | Thyroxine (T4) | T4 | o-(4-hydroxy-3 | 5-diiodophenyl)-3 | 5-diiodotyrosine | thyroxin | thyroxinal | tetraiodothyronine | L-THYROXINE SODIUM | Thyroid hormone | Thyroxine [KLH]

Grado

BioReagent, Suitable for molecular biology

Specifiche e purezza

BioReagent, Suitable for molecular biology

Tipo di molecola

Proteine

Product Line

Prodotti biologici

Stoccaggio e spedizione

Condizioni di conservazione di stoccaggio

Protected from light,Store at -20°C,Avoid repeated freezing and thawing

Spedito in

Ice chest + Ice pads

Stabilità e conservazione

Aliquot and store at -20 °C long term (12 months). Store in the dark. Avoid repeated freeze-and-thaw.

Application Protocols

No item-specific, validated protocols are provided in the Product Data. The following are general, literature-based procedures for T4–protein conjugates. Optimize per application and refer to your antibody supplier’s recommendations.

A. ELISA coating antigen (competitive format):

• Dilute T4–KLH to 1 µg/mL in carbonate buffer (pH 9.6). Add 100 µL/well to high-binding plates and incubate 2 h at 25°C or overnight at 4°C in the dark.
• Wash 3× with PBS-T (0.05% Tween-20). Block with 3% BSA in PBS for 1 h at 25°C.
• Prepare T4 standards in assay buffer and preincubate with primary anti-T4 antibody (per vendor instructions).
• Add mixtures to coated wells; incubate 30–60 min. Wash and add HRP-labeled secondary; develop with TMB; read at 450 nm.

B. Immunogen preparation (research use in animals, per IACUC/ethical guidelines):

• Emulsify T4–KLH with an appropriate adjuvant (e.g., alum, CFA/IFA as permitted). Typical protein doses range widely; determine based on species and protocol. Administer via approved routes and schedules.

C. Dot blot/control:

• Spot 0.1–1 µg of T4–KLH on nitrocellulose, air dry, block with 5% milk, probe with anti-T4 antibody; develop by chemiluminescence.

Note: Concentrations, incubation times, and buffers are starting points only and are not specifications for this item.

Biological Roles

Context (literature/general biology; not a statement of product function in vivo):

• Thyroxine (T4) is an iodinated thyroid hormone synthesized by the thyroid gland of vertebrates. It serves as a prohormone that can be deiodinated to triiodothyronine (T3), the more bioactive form. T4 and T3 regulate gene expression through nuclear thyroid hormone receptors affecting metabolism, development, and thermogenesis.
• Structural features relevant to recognition: The tetraiodinated diphenyl ether core, phenolic hydroxyl, and amino acid side chain define antigenic determinants. Antibodies may differentiate T4 from structurally related iodothyronines based on iodination pattern and orientation of the side chain (literature).
• Carrier protein (KLH): KLH is a potent immunostimulatory carrier widely used to enhance immune recognition of small haptens. It provides T-cell epitopes and promotes robust antibody responses without specific mammalian homologs (reducing tolerance issues in many species; literature observation).

Use in research:

• T4–KLH enables the immune system to recognize the otherwise poorly immunogenic T4 hapten, facilitating the generation and characterization of anti-T4 antibodies for analytical assay development, fundamental endocrinology studies, and method validation.

Note: The product is intended strictly for research use in vitro or in appropriate animal model workflows within institutional guidelines. No medical or diagnostic use is implied.

Buffer Applications

As a protein–hapten conjugate, T4–KLH is commonly handled in standard protein buffers. Item-specific buffer composition is Not specified; the following are general, literature-based practices:

Common buffer systems:

• PBS (10–50 mM phosphate, 150 mM NaCl, pH 7.2–7.4): General handling, dilutions, storage aliquots.
• TBS (20–50 mM Tris, 150 mM NaCl, pH 7.4–8.0): Alternative for some assays; check compatibility with conjugation chemistries.
• Carbonate–bicarbonate (50–100 mM, pH 9.2–9.6): Passive coating of ELISA plates (2–18 h at 4–25°C), followed by blocking.
• Blocking buffers: 1–5% BSA, casein, or fish gelatin in PBS/TBS; include 0.05% Tween-20 for washing steps to minimize nonspecific binding.

Practical tips:

• Sterile-filter working solutions (0.22 µm) and prepare single-use aliquots to prevent contamination and freeze–thaw degradation.
• Avoid chelators (e.g., high EDTA) only if downstream assays require divalent cations; otherwise, low EDTA can help minimize metal-catalyzed oxidation.
• For long-term storage, stabilizers such as 10–50% glycerol (v/v) or 0.2–0.5 M trehalose are commonly used; confirm compatibility with your application.

These buffer guidelines are general. For lot-specific formulation (pH, salts, preservatives) of this item, refer to the CoA/Spec Sheet.

Green Alternatives

Given this product is an aqueous protein–hapten conjugate, green chemistry considerations center on buffer selection, preservative choices, and minimizing waste rather than replacing an organic solvent or hazardous reagent.

Greener practice options (general guidance):

• Buffer systems: Prefer phosphate or bicarbonate buffers prepared from minimal salts; avoid volatile amines where unnecessary. Use CO2-independent buffers only as required.
• Preservatives: Where biosafety policies allow, consider non-azide preservation (e.g., 0.02–0.05% ProClin or sterile single-use aliquots) to reduce heavy metal–azide waste. Verify compatibility with downstream peroxidase-based assays.
• Stabilizers: Use renewable disaccharides (e.g., trehalose, sucrose) or glycerol from bio-based sources for cryo/lyoprotection; avoid high percentages of organic co-solvents.
• Process efficiency: Prepare concentrated stocks and aliquot to minimize freeze–thaw waste. Employ low-bind plastics to reduce protein loss and consequent overuse.

Illustrative comparison (general; not item-specific):

• Sodium azide vs. single-use sterile aliquots
  • Pros (azide): convenient antimicrobial preservation; Cons: toxic, hazardous waste, incompatibility with HRP.
  • Pros (aliquots): eliminates azide, lowers hazard; Cons: requires freezer space, planning.

Conclusion: The most impactful green choices with T4–KLH are sterile technique, right-sized aliquots, and benign buffer systems, reducing both chemical hazards and resource consumption.

Pharmaceutical Uses

This product is supplied for research use only. It is not formulated or qualified for pharmaceutical manufacturing or clinical applications.

General context (literature/general knowledge):

• Hapten–carrier conjugates like T4–KLH are widely used in research settings for immunogen preparation, assay development, and as analytical controls. KLH itself has been explored as a carrier in vaccine research, but such uses require dedicated GMP manufacturing, rigorous characterization (e.g., defined hapten density, residual solvents, endotoxin), and extensive regulatory oversight—none of which are specified for this item.
• Excipients/prescription use: T4–KLH has no role as a pharmaceutical excipient. Any therapeutic or diagnostic application would necessitate separate product design, validation, and regulatory clearance.

If you require materials for translational or manufacturing use, consult vendors offering GMP-grade conjugates with full release testing, validated analytical methods, and change-control documentation.

Physical Properties

Item-specific physical data:

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight (exact): Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: Not specified for this item; refer to CoA/Spec Sheet.
• pH (as supplied): Not specified for this item; refer to CoA/Spec Sheet.
• Extinction coefficient / A280: Not specified for this item; refer to CoA/Spec Sheet.

General/literature expectations for T4–KLH conjugates (not specifications):

• Physical state: Typically supplied as an aqueous solution, suspension, or lyophilized solid; readily reconstituted in neutral physiological buffers (e.g., PBS, pH 7.2–7.4).
• Solubility: KLH and its conjugates are water-soluble; insoluble in most organic solvents. High ionic strength (>0.5 M) and extreme pH (<5 or >9) can reduce solubility (literature).
• Stability: Sensitive to heat, freeze–thaw cycling, and light; iodinated haptens may undergo deiodination/oxidation under harsh conditions. Protect from light (literature), aligning with the item’s storage guidance.
• Aggregation: Protein conjugates can self-associate; gentle handling and inclusion of salt (e.g., 150 mM NaCl) and low concentrations of non-ionic detergents (e.g., 0.01% Tween-20) can minimize nonspecific adsorption/aggregation in working solutions (literature).
• pI/charge: KLH is heterogeneous with multiple isoforms; overall net charge near neutral pH supports good aqueous behavior (literature).

Note: Numerical values such as exact concentration, hapten density, refractive index, or residual salts/buffers are not specified for this item; consult the CoA/Spec Sheet for lot-specific details.

Quality and Grades

Item-specific grade (from Product Data):

• Grade/Purity: BioReagent, Suitable for molecular biology.

What this grade typically implies (general guidance):

• Bioreagent quality denotes preparation suitable for common biochemical workflows (e.g., ELISA, immunization, assay development), with attention to low contaminant levels that could interfere with enzymatic processes or antibody generation. “Suitable for molecular biology” suggests manufacturing controls to limit nucleases, proteases, and microbial contaminants; however, the absence or exact limits of such activities are Not specified for this item; refer to CoA/Spec Sheet.

Considerations specific to hapten–carrier conjugates:

• Heterogeneity: Conjugates inherently display a distribution of hapten densities. Lots may define an average hapten-to-carrier (H/C) ratio; if provided, it will appear on the CoA. This parameter influences immunogenicity and assay binding characteristics.
• Endotoxin/bioburden: For immunization or cell-based assay use, low endotoxin is beneficial. Endotoxin specifications are Not specified for this item; refer to CoA/Spec Sheet.
• Preservatives/stabilizers: Presence or absence of agents such as sodium azide, glycerol, trehalose, or BSA can affect downstream applications. Composition is Not specified for this item; refer to CoA/Spec Sheet.

Recommendation:

• For regulated or highly sensitive applications, request the CoA/Spec Sheet for lot-specific data (protein concentration, H/C ratio, buffer, preservatives, sterility/endotoxin).

Reaction and Applications

Primary research applications (general/literature; expand on manufacturer’s category placement as a protein reagent):

• Immunogen for antibody generation: T4–KLH serves as a robust immunogen to elicit polyclonal or monoclonal antibodies against thyroxine in animal models. The KLH carrier provides T-cell help and enhances immunogenicity of the small T4 hapten.
• Assay development: Utilized as a coating antigen or competitive tracer surrogate in ELISA, CLIA, or LFA development for T4 detection in research matrices. In competitive formats, soluble T4 competes with immobilized T4–protein for antibody binding.
• Positive control/standardization reagent: Provides a consistent, immobilizable T4 epitope density to benchmark assay performance and evaluate anti-T4 antibody specificity and cross-reactivity (e.g., vs. T3, rT3; literature practice).
• Conjugation reference: Serves as a model hapten–protein conjugate for optimizing coupling chemistries, blocking buffers, and surface immobilization protocols.

Practical considerations:

• Hapten density: Higher densities can improve immunogenicity but may reduce antigen accessibility or alter binding kinetics. Optimize empirically.
• Orientation and linker: The conjugation site on T4 and linker length influence antibody specificities (phenolic vs. carboxylate-directed epitopes).
• Plate coating: For ELISA, coat at low µg/mL levels in carbonate buffer (pH ~9.6), then block to reduce nonspecific binding (see Application Protocols). Exact conditions for this item are Not specified; optimize by gradient.

Note: This section reflects general laboratory use cases for T4–KLH conjugates; the product is for research use only.

Reaction Conditions

Because this product is already a thyroxine–KLH conjugate, there are no item-specific reaction conditions to create it provided here. However, for users optimizing downstream assay steps or preparing analogous materials, the following literature-based conditions are commonly employed (general guidance; not item-specific specifications):

• ELISA plate coating with T4–protein conjugates:

  • Buffer: 50–100 mM carbonate–bicarbonate, pH 9.2–9.6.
  • Concentration: 0.5–5 µg/mL conjugate; 100 µL/well.
  • Incubation: 2 h at 25°C or overnight at 4°C.
  • Post-coating: Block with 1–5% BSA or casein in PBS/TBS for 1 h at 25°C.

• Competitive assay setup:

  • Prepare serial dilutions of free T4 standard in assay buffer.
  • Incubate anti-T4 antibody with standards/samples, then transfer to plates coated with T4–protein.
  • Detect with species-appropriate secondary antibodies (HRP/AP or fluorescent), adhering to supplier guidelines.

• Conjugation (for users preparing new T4–KLH lots):

  • Activation: EDC/NHS activation of T4-carboxylate in MES buffer (50 mM, pH 5.5–6.0), followed by coupling to KLH in PBS (pH 7.2–7.4).
  • Typical molar ratios and times vary; optimize to achieve desired hapten density while preserving protein solubility (monitor by UV/Vis or mass spectrometry where applicable).

For this product, exact coating concentrations, buffer composition, or hapten density are Not specified; consult the CoA/Spec Sheet and optimize empirically for your assay.

Safety and Handling

GHS/Classification (from Product Data):

• Signal word: Not specified for this item; refer to SDS.
• Hazard statements (H-phrases): Not specified for this item; refer to SDS.
• GHS classification and pictograms: Not specified for this item; refer to SDS.

General laboratory safety guidance (literature/practice):

• PPE: Wear lab coat, nitrile gloves, and safety glasses when handling protein conjugates and buffers. Handle solutions in a clean biosafety cabinet if sterility must be maintained.
• Allergenicity/sensitization: Carrier proteins (including KLH) may present sensitization risks by skin or inhalation of aerosols/lyophilized powder. Avoid generating dust or aerosols; use containment and appropriate respiratory protection if aerosolization is possible (literature).
• Light sensitivity: Protect from light to prevent photodegradation of iodinated haptens (consistent with Product Data).
• Incompatibilities: Avoid strong oxidizers, extremes of pH, and organic solvents that denature proteins. Do not mix with sodium azide if downstream HRP-based assays are planned (azide inhibits HRP), unless compatible with the intended use.
• First aid (overview): In case of skin/eye contact, rinse with water for 15 minutes. If inhaled, move to fresh air. If ingested, rinse mouth with water. Seek medical attention as appropriate. Defer to the SDS for authoritative instructions.
• Waste: Dispose of protein-containing waste according to institutional biosafety and chemical waste policies.

Always consult the Safety Data Sheet (SDS) for definitive hazard, exposure controls, and spill/first-aid procedures.

Solvent Selection

This product is a protein–hapten conjugate and is not used with organic solvent systems in the way small molecules are. Selection focuses on aqueous buffers that maintain protein stability and functional presentation of the T4 epitope.

General buffer guidance (literature/general):

• Primary buffer: Phosphate-buffered saline (PBS), pH 7.2–7.4, 10–50 mM phosphate, 150 mM NaCl, supports solubility and isotonicity for most immunochemical workflows.
• Alternative buffers: Tris-buffered saline (TBS, 20–50 mM Tris, 150 mM NaCl, pH 7.4–8.0) is acceptable for many assays but may interact with some crosslinkers or metal chelates.
• Coating buffers (ELISA): Carbonate–bicarbonate, pH 9.2–9.6, can improve passive adsorption to polystyrene plates (use for coating; switch to PBS/TBS for blocking and assays).
• Additives: 0.01–0.05% Tween-20 reduces nonspecific adsorption; 0.1–1% BSA or casein as blockers in assays; 5–50% glycerol or 0.2–0.5 M trehalose for storage stabilization (as appropriate). Composition for this item is Not specified; verify compatibility.
• Organic solvents: Avoid (>5% v/v) as they can denature KLH; if needed for specific surface-modification workflows, keep below denaturing thresholds and confirm stability empirically.

Practical tips:

• Filter-sterilize working buffers (0.22 µm) to limit bioburden.
• Maintain neutral pH and moderate ionic strength to minimize aggregation and preserve epitope presentation.

Storage and Reconstitution

Item-specific storage (from Product Data):

• Storage conditions: Protect from light; store at −20°C; avoid repeated freezing and thawing.
• Shipment: Ice chest + ice pads.

Item-specific format/reconstitution details: Not specified for this item; refer to CoA/Spec Sheet.

General guidance (literature/practice):

• If lyophilized: Reconstitute with sterile, degassed PBS (pH 7.2–7.4) to the desired concentration. Allow to dissolve gently on ice without vigorous vortexing.
• If supplied in solution: Upon receipt, mix by gentle inversion, aliquot into low-binding tubes to single-use portions, and freeze at −20°C or −80°C as appropriate.
• Stabilizers: For frozen storage, consider adding 10–50% glycerol (v/v) to permit storage at −20°C without freezing, or include 0.2–0.5 M trehalose for lyophilized stocks. Verify compatibility with downstream assays.
• Light protection: Wrap vials in foil or store in amber containers to protect the iodinated hapten.
• Thawing: Thaw on ice. Do not refreeze the same aliquot; repeated freeze–thaw can cause aggregation and loss of activity.
• Sterility: If sterility is required, work aseptically and consider 0.22 µm filtration of reconstituted solutions (note potential protein loss on membranes; pre-rinse filters).

Always consult the lot-specific CoA/Spec Sheet for exact formulation, concentration, and any preservatives present.

Structure and Identity

Overview: Thyroxine–KLH (T4–KLH) is a hapten–carrier conjugate in which the small-molecule hormone thyroxine (T4, L-3,5,3′,5′-tetraiodothyronine) is covalently attached to the high–molecular-weight carrier protein keyhole limpet hemocyanin (KLH). This produces a heterogeneous mixture of protein molecules bearing multiple T4 moieties.

Item-specific identifiers (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 / InChIKey / CAS / CID: Not specified for this item; refer to CoA/Spec Sheet.

General structural features (literature/general knowledge):

• Carrier protein: KLH is a large, copper-containing, oxygen-transport protein from Megathura crenulata; native oligomers are megadalton-scale and typically used in subunits for conjugation.
• Hapten: Thyroxine (T4) contains a diphenyl ether core with four iodine atoms and a zwitterionic amino acid side chain (phenolic and carboxylate functionalities). In conjugates, T4 is usually derivatized (e.g., via the carboxylate) and linked to lysine side chains on KLH through amide bonds using carbodiimide or heterobifunctional linkers (literature).
• 2D description in words: A large globular protein surface decorated with multiple pendant T4 groups attached via short organic linkers to exposed ε-amine groups of lysine residues. Each T4 moiety presents an iodinated diphenyl ether ring system with phenolic oxygen and an alanine-like side chain.

Notes on identity:

• Conjugation yields a distribution of hapten densities (moles T4 per mole of KLH), not a single stoichiometric entity (literature). Any stated hapten-to-carrier ratio, if controlled, would appear on the CoA.

Synthetic Utility

This item is a finished hapten–protein conjugate rather than a small-molecule building block; it is not typically used as a substrate or reagent in organic synthesis. Nevertheless, it has utility in synthetic/analytical methodology development within bioconjugation and immunochemistry (general literature):

• Conjugation benchmarking: Use T4–KLH as a model to evaluate coupling strategies (e.g., EDC/NHS amide formation, maleimide–thiol coupling with derivatized T4, or click chemistry with azido/alkyne-modified haptens). Although the present item is already conjugated, parallel preparations help optimize linker chemistry and hapten density.
• Surface immobilization: Serves as a test analyte for immobilization protocols on ELISA plates, sensor chips (SPR), and microarrays, informing surface activation (e.g., NHS-ester, epoxide) and blocking strategies.
• Antibody screening/selection: Facilitates panning and selection of binders (hybridoma screening, phage display) by presenting the T4 epitope in a multivalent format.
• Analytical method validation: Aids in establishing calibration curves and competitive binding windows when paired with free T4 standards and anti-T4 antibodies.

Note: If your objective is chemical synthesis of T4 derivatives or bespoke conjugates, procure unconjugated thyroxine and KLH (or alternative carriers such as BSA/OVA) and apply controlled conjugation methods. The current product’s hapten density and composition are Not specified; refer to CoA for any available characterization.

Target Specificity

Item-specific target information from Product Data: Not specified for this item; refer to CoA/Spec Sheet.

General context (literature):

• The conjugate presents thyroxine (T4) epitopes on a KLH carrier. In immunochemical workflows, anti-T4 antibodies raised or tested with T4–KLH may display specificity for T4 over related iodothyronines (e.g., T3, rT3), but actual specificity and cross-reactivity profiles depend on antibody clone and conjugation chemistry. Such parameters are determined empirically and are not defined for this item.