Dual Color SDS-PAGE Protein Sample Loading Buffer (5×) is an independently developed product by Aladdin. It is an improved 5-fold concentrated protein loading buffer that is safer and odor-free, containing two dyes: blue and red. The migration rate of the blue dye matches that of bromophenol blue, while the red dye migrates slightly faster than the blue dye. For gels with concentration higher than 12%, the red dye migrates a little slower than the blue dye. The red dye can be transferred together with proteins onto PVDF or NC membranes, which allows lane tracing after membrane transfer. This product thaws rapidly after removal from -20°C storage. It is non-viscous and saves operation time. A new odorless and more stable reducing agent is adopted to replace DTT and β-mercaptoethanol. It is suitable for sample loading in SDS-PAGE. Its performance is consistent with conventional SDS-PAGE protein loading buffer, with no significant differences observed.
Instructions for Use:
1. Thaw completely at room temperature (10~30°C) and mix well. Note: For faster preparation, thaw in a 37°C water bath or metal bath. Minimize the incubation time in water bath or metal bath after thawing.
2. Add 5 μL loading buffer to every 20 μL protein sample (5-fold dilution), and mix thoroughly. Note: If the protein sample is overly concentrated, dilute the sample with double-distilled water before adding loading buffer.
3. Heat the mixture in a 100°C water bath for 5–10 min to denature proteins.
4. Cool down to room temperature, centrifuge at 10000–14000 rpm for 2–5 min, then take the supernatant for direct loading.
5. Normally stop electrophoresis when bromophenol blue migrates to approximately 1 cm above the bottom of the gel.
Precautions:
1. Due to different reducing agents, the odor-free version and DTT-containing version of SDS-PAGE protein loading buffer may yield different electrophoresis results for individual proteins.
2. This product contains denaturants and is not applicable to Native-PAGE.
3. The product must be fully thawed and homogenized prior to use.
4. For your safety and health, wear a lab coat and disposable gloves.
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