Dual Color SDS-PAGE Protein Sample Loading Buffer

Cat. No.: D752143

GRADE & PURITY 6X, Odorless

Overview

Aladdin's Dual Color SDS-PAGE Protein Sample Loading Buffer (6X, Odorless) is an improved protein loading buffer with red and blue dyes, which can be used for the preparation and loading of protein samples onto a gel for SDS-PAGE analysis.This product is safe to use. Instead of odoriferous DTT or 2-Mercaptoethanol, this product employs an odorless and more stable reducing agent with similar reducing capability.In conventional SDS-PAGE, the blue dye migrates exactly the same as bromophenol blue, and the red dye migrates similarly to the blue dye. In Tris-Gly electrophoresis system, the red dye migrates slightly faster than the blue dye in SDS-PAGE gel lower than 15%. While in SDS-PAGE higher than 15%, the red dye migrates slightly slower than the blue dye.This product can be used for lane tracing after membrane transfer. It contains a special red dye which can be transferred to a PVDF or NC membrane to indicate the position of the lane, facilitating operations such as film cutting. Please refer to Figure 1 for the electrophoresis and the transfer results of this product.Figure 1. Aladdin's Dual Color SDS-PAGE Protein Sample Loading Buffer (6X, Odorless) in the UltraBio™ Plus PAGE Precast Gel (Tris-Gly, 12%, 10 wells) after electrophoresis and transfer, as well as in the PVDF membrane after transfer (Blot).

Precautions：

For denaturing PAGE analysis, protein samples mixed with this product is recommended to be heated at 95℃ for 5 minutes. Too high a temperature (e.g., 100℃) or too long a time (e.g., more than 15 minutes) may result in protein degradation or abnormal color change of the indicator.This product must be completely dissolved and mixed well before use.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.

Instructions for Use：

1. Thaw this product at room temperature or in a water bath no more than 37℃. Place at room temperature immediately once dissolved and store at -20℃ after use.2. Add 1μl of this product per 5μl of protein sample, and mix well. Note: After dilution to 1X, this product can also be used for sample lysis directly.3. Heat at 95℃ for 5-10 minutes to fully denature proteins.Note: If the starting material is large and the genomic DNA content is high, the mixture may be very sticky after boiling. In this case, heat for another 5-10 minutes directly or after addition of more 1X sample loading buffer. The thorough heating will release proteins bound to genomic DNA and break genomic DNA so that the mixture will be not too sticky for sample loading. Appropriate ultrasound or repeated aspiration with a 1ml pipette can also help.4. After cooling to room temperature, centrifuge briefly at room temperature, then load the supernatant onto a SDS-PAGE gel.5. Stop electrophoresis when the red or blue dye reaches the bottom of the gel.

Specifications

Product Name

Dual Color SDS-PAGE Protein Sample Loading Buffer

Specifications & Purity

6X, Odorless

Molecule Type

Bioreagents/Buffer

Product Line

biological_products

Storage and Shipping

Concentration

6X, Odorless

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Stability And Storage

Stored at -20 ℃, valid for at least one year.

Application Protocols

General SDS-PAGE sample preparation protocol (literature/general; adapt to your gel system and consult SKU D752143 CoA/Spec Sheet for any product-specific details):

1. Thaw buffer on ice. Mix gently to ensure homogeneity. Avoid introducing bubbles.
2. Combine sample with loading buffer to achieve 1× final concentration. If needed, add reducing agent (if not already included) to the desired final level.
3. Incubate for denaturation: 95–100°C for 3–10 min (or 70°C for 10–20 min for heat-sensitive proteins). Cool to room temperature, then briefly centrifuge.
4. Load appropriate volume into gel wells. Use the dual-color dyes to monitor the ion front and mid-range progress.
5. Run electrophoresis under your standard conditions. Stop the run when the faster dye approaches the gel bottom while the slower dye remains in-frame for your targets.
6. Proceed to staining or transfer. If transferring, use dye fronts to standardize transfer start/stop times between gels.

Notes:

• For non-reducing SDS-PAGE, omit reducing agents and avoid excessive heating that could promote artificial aggregation.
• If sample contains high salts, dilute to mitigate conductivity-related band distortions.

Tested application claims and recommended dilutions specific to this item: Not specified for this item; refer to CoA/Spec Sheet.

Biological Roles

Not applicable as a biological molecule with intrinsic physiological function; this is a laboratory formulation used to denature proteins and aid electrophoretic separation.

Contextual notes (literature/general):

• SDS disrupts noncovalent interactions, producing linearized polypeptide chains; this is a laboratory effect rather than a native biological role.
• Reducing agents (when present) cleave disulfide bonds, allowing analysis of subunit composition versus disulfide-linked oligomers.
• Tracking dyes provide visual fronts; they are analytical aids with no biological activity in this context.

No clinical or in vivo uses are implied or recommended; this product is for research use only.

Buffer Applications

Highly relevant. Dual Color SDS-PAGE Protein Sample Loading Buffer is used to prepare protein samples for polyacrylamide gel electrophoresis under denaturing conditions.

Typical use (literature/general; verify product-specific instructions on CoA/Spec Sheet):

• Choose the correct concentrate (e.g., 2×, 4×, 5×). Mix with sample to reach 1× final buffer in the loaded sample.
• If reducing conditions are required and compatible with your target proteins, ensure a reducing formulation or add a reducing agent separately to the desired final concentration.
• Heat samples (commonly 95–100°C for 3–10 min; or 70°C for heat-labile proteins) to facilitate denaturation. Cool, then briefly spin down.
• Load appropriate volume based on gel well capacity and desired total protein (e.g., 1–30 µg per lane for Coomassie staining; adjust for detection method).
• Use the dual-color dye fronts to monitor electrophoresis progression and to time gel termination or transfer initiation consistently.

Practical notes:

• High-salt or high-detergent sample matrices may affect protein mobility; dilute with water or sample buffer to reduce artifacts.
• For native PAGE or non-denaturing studies, do not use SDS-PAGE loading buffer; choose native loading buffers instead.
• For downstream Western transfer, the dye fronts can help prevent over-running the gel and improve blot-to-blot reproducibility.

Green Alternatives

SDS-PAGE loading buffers inherently contain components (e.g., SDS, reducing agents, dyes) that raise EHS considerations. While the function dictates many components, greener practices can mitigate impact.

Greener practice options (literature/general):

• Prefer odourless, low-toxicity reducing agents (e.g., TCEP) over β-mercaptoethanol when compatible with your workflow to reduce VOCs and odors.
• Minimize waste: Prepare only required aliquots and avoid repeated freeze–thaw that leads to disposal.
• Closed handling: Use screw-cap tubes and fume hoods for thiol-containing formulations to limit emissions.
• Aqueous cleanup: Rinse glassware promptly to prevent SDS residues that require excessive detergent/solvent for removal.
• Energy-efficient runs: Optimize gel percentage and run voltage to reduce time and power consumption.

Comparison (general):

• Conventional reducing agent (β-mercaptoethanol): Strong odor, volatile, hazardous to aquatic life; effective disulfide reduction.
• Alternative (TCEP): Odorless, non-volatile, stable in air; may interact differently with certain labeling chemistries.

Item-specific composition and available greener variants for SKU D752143: Not specified for this item; refer to CoA/Spec Sheet.

Pharmaceutical Uses

Not a pharmaceutical or excipient product. This item is intended for laboratory research workflows only.

Industry-relevant context (general):

• In biopharmaceutical development and QC labs, SDS-PAGE loading buffers are routinely used for in-process characterization of proteins (e.g., titer, purity checks, aggregation/fragmentation under reducing/non-reducing conditions).
• For GMP-regulated environments, buffer composition, lot traceability, and qualification are critical; obtain and retain the CoA/Spec Sheet and SDS. Confirm whether the formulation meets your site’s qualification criteria.

No therapeutic, diagnostic, or clinical use is claimed or supported. For research use only.

Physical Properties

• Physical state/appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration (e.g., 2×, 4×, 5×): Not specified for this item; refer to CoA/Spec Sheet.
• pH: Not specified for this item; refer to CoA/Spec Sheet.
• Density, refractive index, viscosity: Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: Aqueous formulation (mixture); miscible with water (general/literature for SDS-PAGE buffers).

Contextual notes (literature/general):

• Common formulations are water-based and readily mix with protein samples in aqueous buffers, urea/thiourea solutions, or sample lysis buffers.
• Typical pH for Laemmli-type systems is near neutral to slightly basic (around pH 6.8 for stacking buffer context; pH is formulation dependent, literature).
• Color: “Dual color” denotes inclusion of two tracking dyes for visual front and mid-range migration; exact dye identities and intensities vary by manufacturer.

Do not treat literature values as product specifications. For exact physical attributes and handling characteristics of SKU D752143, consult the product’s CoA/Spec Sheet.

Quality & Grades

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

Context and expectations for this product type (general):

• Ready-to-use loading buffers are typically produced to consistent formulation and appearance standards to ensure reproducible protein denaturation, dye migration, and lane loading density.
• Dual color indicates inclusion of two tracking dyes to monitor electrophoresis progress across a broad molecular-weight range; this aids in run timing and gel-to-gel consistency.
• For high-sensitivity downstream applications (e.g., silver staining, fluorescent stains, or MS), low-background formulations and clean raw materials are preferred; consult the CoA for details relevant to your workflow.
• Lot-specific QC may include visual inspection (clarity/color), pH verification, and functional checks (e.g., predictable tracking dye mobility). Exact QC tests for SKU D752143 are Not specified for this item; refer to CoA/Spec Sheet.

If your application imposes special constraints (e.g., thiol-free for alkylation studies, amine-free for labeling chemistries, or MS-compatibility), verify composition and grade on the CoA/Spec Sheet before use.

Reaction & Applications

This product is not used as a chemical reagent for synthesis; it is used for preparing protein samples for SDS-PAGE.

Applications and practical notes (literature/general):

• Protein denaturation and uniform charge: SDS binds to proteins, unfolding them and imparting a near-constant charge-to-mass ratio, enabling size-based separation.
• Disulfide reduction (if reducing agent present): Breaking disulfide bonds yields fully denatured subunits for accurate apparent molecular weight estimation.
• Tracking dyes (dual color): Provide two electrophoretic fronts to monitor progress in real time and standardize stopping points across gels.
• Density agent (e.g., glycerol): Increases sample density for reliable well loading and minimal sample loss.

Practical tips:

• Mix sample with the loading buffer at the manufacturer-recommended ratio (e.g., 1:1 with 2×; 1:3 with 4×; 1:4 with 5× – ratios are literature examples only). Exact ratio for this item is Not specified for this item; refer to CoA/Spec Sheet.
• Heat treatment (commonly 95–100°C for 3–10 min, or 70°C for sensitive proteins) is a literature-standard practice; verify suitability for your protein and confirm any product-specific guidance on the CoA/Spec Sheet.
• Briefly spin samples to collect condensate and prevent smiling or crooked lanes due to uneven loading.

Reaction Conditions

Not a chemical reaction reagent. However, common handling conditions for SDS-PAGE sample preparation (literature/general) include:

• Mixing ratio: Adjust to reach 1× final loading buffer in the sample (e.g., 1:1 with 2×; 1:3 with 4×; 1:4 with 5×). Exact ratio for SKU D752143 is Not specified for this item; refer to CoA/Spec Sheet.
• Temperature: Heat denaturation commonly at 95–100°C for 3–10 minutes; alternatives include 70°C for 10–20 minutes for sensitive proteins.
• Atmosphere: No special atmosphere required; avoid aerosol generation. If using thiols, work in a fume hood.
• Timing: Process samples promptly after mixing/heating to prevent precipitation or re-aggregation.
• Additives: Protease inhibitors and alkylating agents (e.g., iodoacetamide post-reduction) may be included in upstream steps; avoid introducing reagents that react with dyes or SDS.

For precise, product-specific instructions (e.g., recommended heating protocol, compatibility notes), consult the CoA/Spec Sheet.

Safety & Handling

Authoritative safety information is provided in the SDS for this specific product. The following are general considerations for SDS-PAGE loading buffers.

• GHS classification, signal word, pictograms, H-statements: Not specified for this item; refer to SDS.
• Primary hazards (literature/general):
  • May contain SDS, an anionic surfactant that can cause skin/eye irritation and is harmful if inhaled as aerosol.
  • May contain reducing agents (e.g., DTT, β-mercaptoethanol, TCEP) that can be irritant; β-mercaptoethanol is strongly odorous and toxic—use in a fume hood.
  • Dyes and buffers may stain; avoid contact with skin and clothing.
• PPE: Wear lab coat, safety glasses, and appropriate chemically resistant gloves. Avoid inhalation of vapors/aerosols. Handle potential thiols in a fume hood.
• Incompatibilities (general): Strong oxidizers (if reducing agents are present). Avoid contact with acids/bases that could decompose buffer components.
• First aid (general guidance; defer to SDS):
  • Skin/eye contact: Rinse with water for several minutes; remove contaminated clothing; seek medical advice if irritation persists.
  • Ingestion/inhalation: Move to fresh air; rinse mouth; seek medical attention if unwell.
• Spill response: Absorb with inert material, collect for disposal. Clean surfaces with water and detergent.
• Waste: Dispose according to institutional and local regulations for laboratory chemical waste.

Always review and follow the product’s SDS and institutional EHS practices.

Solvent Selection

This item is an aqueous electrophoresis loading buffer; external solvent selection is typically not required.

• Matrix: Water-based mixture formulated for direct combination with protein samples.
• Compatibility: Mixes with common biological buffers (e.g., PBS, Tris, HEPES) and denaturants (urea/thiourea) used in protein sample prep (literature/general). Avoid excessive organic cosolvents that can precipitate SDS or distort dye mobility.
• When to choose: Use a dual-color buffer when visual monitoring of the electrophoresis front and mid-range mobility is desired, improving timing for stopping the run and transferring.
• Alternatives: Monochrome (single-dye) or dye-free buffers may be preferable for certain imaging modalities or dye-interference-sensitive workflows. Reducing-agent-free versions are used when disulfide status must be preserved prior to targeted reduction.

For exact compatibility claims and any solvent limitations of SKU D752143, consult the CoA/Spec Sheet.

Storage & Reconstitution

• Storage conditions (item-specific): Store at -20°C.
• Shipping (item-specific): Shipped in ice chest + ice pads.
• Appearance/format and reconstitution: Not specified for this item; refer to CoA/Spec Sheet. If provided frozen, thaw on ice and mix gently to ensure uniformity. Avoid introducing foam.
• Aliquoting: To minimize freeze–thaw cycles, prepare single-use aliquots upon first thaw (general best practice for SDS-containing buffers).
• Stability: Not specified for this item; refer to CoA/Spec Sheet. As general guidance, repeated freeze–thaw can lead to precipitation or dye intensity changes; inspect before use.
• Handling: Warm to room temperature only as needed, then return promptly to -20°C storage. If precipitate is observed after thawing, mix gently and allow to fully dissolve before use.

Always consult the product’s CoA/Spec Sheet and SDS for definitive storage and handling guidance for SKU D752143.

Structure & Identity

Dual Color SDS-PAGE Protein Sample Loading Buffer is a formulated mixture (not a single compound) designed for denaturing protein samples and providing visible tracking dyes during electrophoresis.

• Item type: Formulated aqueous buffer (mixture) for SDS-PAGE sample preparation
• SKU: D752143
• Catalog category: Protein reagents (全部 / 可售 / 蛋白质)
• CAS, CID, InChIKey, SMILES: Not applicable to mixtures
• Molecular formula and molecular weight: Not applicable to mixtures

Structural/Compositional features (literature/general):

• Typical SDS-PAGE loading buffers (e.g., Laemmli-type) contain: an anionic surfactant (SDS) for denaturation, buffering salts (usually Tris/Tris-HCl), a polyol (glycerol) for density, one or more tracking dyes (e.g., bromophenol blue and xylene cyanol for “dual color”), and often a reducing agent (e.g., DTT, TCEP, or β-mercaptoethanol) to break disulfide bonds.
• 2D description: aqueous solution in which SDS micelles bind to polypeptides imparting uniform negative charge per mass; tracking dyes migrate at characteristic mobilities for run monitoring.

Item-specific composition for this product (concentrations, exact components): Not specified for this item; refer to CoA/Spec Sheet.

Synthetic Utility

Not applicable. This product is not a building block or reagent for chemical synthesis.

Guidance:

• If your workflow requires chemical modification of proteins (e.g., labeling, crosslinking) prior to SDS-PAGE, perform such reactions before adding SDS-PAGE loading buffer, as its components (anionic surfactant, dyes, reducing agents) can interfere with many coupling chemistries.
• For mass spectrometry or proteomics sample prep, consider MS-compatible buffers and perform detergent removal as required. Composition of SKU D752143 is Not specified for this item; refer to CoA/Spec Sheet.

Target Specificity

Not applicable. This product is a general-purpose protein sample loading buffer and does not possess target specificity (no antigen, clone, isotype, or species selectivity).