Description

5× Protein Loading Buffer (Reducing) is a concentrated SDS-PAGE protein sample loading buffer designed for efficient protein denaturation and reduction before gel electrophoresis. During protein sample preparation, incomplete protein unfolding or insufficient disulfide bond reduction can affect protein migration, molecular weight estimation, and downstream Western blot analysis. This reducing protein loading buffer provides a reliable solution for preparing consistent protein samples by combining strong denaturing conditions with an advanced reducing system.

Unlike traditional SDS-PAGE sample buffers containing DTT or β-mercaptoethanol, this 5× reducing sample buffer uses a new-generation odor-free reducing agent that provides strong reduction capability with improved thermal stability. The optimized formulation maintains reducing performance during high-temperature sample treatment while avoiding the unpleasant odor associated with commonly used reducing reagents.

5× Protein Loading Buffer (Reducing) is suitable for protein electrophoresis workflows including SDS-PAGE, Western blot sample preparation, recombinant protein analysis, and cell or tissue lysate evaluation. The formulation is designed to maintain a neutral pH after heating, helping minimize protein degradation during sample preparation.

Specifications

Product Name	5× Protein Loading Buffer (Reducing)
SKU	AP0181
Product Type	5× SDS-PAGE Protein Sample Loading Buffer
Format	5× Concentrated Reducing Loading Buffer
Volume	1 mL × 10
Reducing System	Odor-free reducing agent, DTT and β-mercaptoethanol free
Sample Ratio	4 μL protein sample + 1 μL 5× loading buffer
Protein Denaturation	100°C heating for 5–10 minutes
Applications	SDS-PAGE, Western blot, protein electrophoresis, protein sample preparation
Storage	-20°C
Shelf Life	24 months

Features

Strong reducing performance: Provides efficient disulfide bond reduction to promote complete protein denaturation before SDS-PAGE analysis.
Odor-free reducing formulation: Eliminates the strong odor associated with β-mercaptoethanol and provides a more convenient laboratory workflow.
High thermal stability: Maintains reducing activity during high-temperature protein sample treatment.
Compatible with SDS-PAGE and Western blot workflows: Supports routine protein electrophoresis and immunoblotting applications.
Optimized sample preparation: Maintains neutral pH after heating to help reduce protein degradation during denaturation.
5× concentrated format: Provides flexible sample preparation with reduced reagent consumption compared with ready-to-use loading buffers.

Applications

5× Protein Loading Buffer (Reducing) is widely used for preparing protein samples in molecular biology, biochemistry, and protein analysis workflows.

SDS-PAGE Protein Electrophoresis: Prepares denatured and reduced protein samples for separation according to molecular weight.
Western Blot Sample Preparation: Generates consistent protein lysates for electrophoresis followed by antibody-based detection.
Cell Lysate Protein Analysis: Suitable for preparing whole-cell protein samples from cultured cells.
Tissue Protein Extraction Analysis: Supports analysis of proteins extracted from biological tissue samples.
Recombinant Protein Characterization: Used for evaluating recombinant protein expression and purity after purification.
Protein Molecular Weight Determination: Enables accurate comparison of protein migration patterns during SDS-PAGE.
Protein Expression Studies: Suitable for research involving protein abundance comparison and expression analysis.
Proteomics Sample Preparation: Provides standardized protein denaturation and reduction before electrophoretic analysis.

Protocol Overview

For SDS-PAGE sample preparation, combine protein samples with 5× Protein Loading Buffer (Reducing) according to the recommended dilution ratio.

Add 1 μL of 5× Protein Loading Buffer to every 4 μL of protein sample.
Mix thoroughly to ensure complete sample homogenization.
Heat samples at 100°C for 5–10 minutes to denature proteins.
Cool samples to room temperature.
Centrifuge at 10,000–14,000 rpm for 5 minutes.
Transfer the supernatant and load directly onto SDS-PAGE gels.

This product contains denaturing components and is intended for SDS-PAGE workflows. It is not suitable for native PAGE or non-denaturing electrophoresis.

Ordering Information

SKU	Product	Size
AP0181	5× Protein Loading Buffer (Reducing)	1 mL × 10

FAQ

What is 5× Protein Loading Buffer (Reducing) used for?

5× Protein Loading Buffer (Reducing) is used for SDS-PAGE protein sample preparation. It denatures proteins and reduces disulfide bonds before electrophoresis, helping generate consistent protein migration patterns for protein analysis and Western blot workflows.

What is the difference between reducing and non-reducing protein loading buffer?

Reducing protein loading buffer contains reducing components that break disulfide bonds within and between proteins. Non-reducing sample buffers do not disrupt these bonds and are used when maintaining disulfide-linked protein structures is required.

Can this reducing sample buffer be used for Western blot?

Yes. This SDS-PAGE reducing sample buffer is suitable for preparing protein samples before electrophoresis and Western blot detection.

Does this protein loading buffer contain DTT or β-mercaptoethanol?

No. This odor-free reducing protein loading buffer does not contain DTT or β-mercaptoethanol. It uses an alternative reducing system designed to provide strong reduction performance with improved laboratory handling.

Why use a 5× protein loading buffer instead of a 2× sample buffer?

A 5× concentrated loading buffer allows flexible sample preparation with a smaller buffer volume while maintaining effective protein denaturation and reduction conditions.

Can this buffer be used for native PAGE?

No. Because this product contains denaturing components, it is designed for SDS-PAGE and is not suitable for native PAGE or non-denaturing electrophoresis.

How should 5× Protein Loading Buffer (Reducing) be stored?

The product should be stored at -20°C. Under recommended storage conditions, the shelf life is 24 months.