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Cat. No.: QCSP-1 (for 1ml)

Description

QuickColor™ SDS-PAGE Protein Staining and Loading Buffer (5X) is a special SDS-PAGE protein loading buffer that contains a safe and non-toxic fluorescent dye. It allows proteins to be stained purple and exhibit orange fluorescence when boiled with the loading buffer prior to sample loading. This enables direct visual observation of the purple protein bands during or after electrophoresis, or the orange fluorescence bands using UV or blue light equipment.

This product utilizes safe and non-toxic fluorescent dyes, eliminating the need for additional staining and destaining steps. The staining step is completed during the boiling process of proteins with the loading buffer, allowing for direct observation and recording of the protein electrophoretic patterns after electrophoresis. It is compatible with UV and blue light detection systems, as well as subsequent Western blotting procedures.

Features

• Safe and Convenient Staining: This product uses safe and non-toxic dyes and does not require any additional experimental steps. Simply follow the standard protocol for SDS-PAGE protein loading buffer (5X). Add 1/4 volume of this product to the protein sample, mix well, and heat in a 95-100°C water bath for 5 minutes. This will result in protein staining, turning the protein purple while imparting orange fluorescence. Staining and electrophoresis can be performed under normal lighting conditions without the need for additional light protection measures.
• No Impact on Electrophoretic Mobility: The small size and neutral charge of the dye molecules do not affect the electrophoretic mobility of protein bands or the final electrophoretic patterns.
• Immediate Observation of Sample Electrophoresis: Using this product, the electrophoretic results of protein samples can be observed immediately after electrophoresis. Protein bands with quantities larger than 500 ng can be visually observed as purple bands, while for samples with lower protein quantities, fluorescence can be observed using UV or blue light (requires the use of a blue light filter) in a darkroom.
• No Gel Staining Required: After using this product, there is no need for any gel staining procedures. Protein samples can be directly photographed to record the electrophoretic patterns. For samples with high protein quantities, direct photography is possible, and for samples with lower protein quantities, UV or blue light imaging equipment can be used for photography and documentation.
• Compatible with Western Detection: After using this product for sample loading and electrophoresis, direct transfer and subsequent Western detection can be performed without any additional steps. PVDF membranes and nitrocellulose membranes (NC membranes) are both compatible. After transfer, protein bands can be observed as purple bands or orange fluorescence bands. Testing with various commonly used antibodies has shown that the use of this product does not significantly affect the sensitivity of Western detection. Similar products from abroad are also perfectly compatible with Western detection. However, it cannot be ruled out that this product may affect the detection of specific antibodies for target proteins.
• Low Background, Free Dye Does Not Affect Observation and Detection: Free dye molecules migrate at a similar rate to the tracking dye bromophenol blue, and they collectively migrate to the end of the gel, ensuring a clean and transparent background.
• High Sensitivity, as low as 25 ng: After using this product, protein bands modified by fluorescent dyes appear as pinkish-purple bands, visible to the naked eye under daylight conditions for single protein bands of 0.5 μg or above (Figure 1). Protein bands of 0.2 μg are also faintly visible. Using UV or blue light imaging equipment, protein bands as low as 25 ng can be detected (Figure 2).
• Compatible with UV and Blue Light Imaging Systems: Whether using UV imaging equipment or safe blue light imaging devices, protein bands as low as 25 ng can be detected.
• Compatible with Tris-glycine and Hepes Gel Systems: It is compatible with the most common Tris-glycine gel system as well as the less common Hepes gel system.
• Good Staining Stability: After SDS-PAGE electrophoresis, the fluorescence of protein bands remains stable for at least 5 hours when stored at 4°C. If immediate transfer is performed after electrophoresis, the fluorescence of protein bands on the membrane can be stable for at least 24 hours when stored at 4°C.

Application

This product is primarily used to directly observe, record, and confirm the equal loading of each lane during Western detection, which is particularly important for Western detection of tissue samples. It also allows for rapid and convenient observation of the induced expression and separation/purification effects of target proteins during SDS-PAGE.

Precautions

• Low pH significantly affects the staining results. If the pH of the protein sample solution is below 5, it is recommended to adjust the pH to 7 or above for better staining results.
• Most pre-stained and non-stained protein molecular weight markers are not compatible with this product. However, non-stained protein molecular weight markers dissolved in water or physiological saline without the addition of loading buffer can react with QuickColor™ SDS-PAGE Protein Staining and Loading Buffer (5X) to generate protein molecular weight markers with orange fluorescence.
• Protein bands stained with this product are not recommended for subsequent protein sequencing, protein mass spectrometry analysis, or antibody preparation.
• Store this product at -20°C, protected from light. If stored at room temperature for an extended period (more than 2-3 weeks), the staining efficiency may decrease due to oxidation. In such cases, adding an appropriate amount of DTT can restore the clarity and brightness of the protein bands. However, the final concentration of DTT should not exceed 20 mM.
• This product is harmful to humans. Handle with care and take necessary precautions to avoid direct contact or inhalation.
• This product is intended for scientific research purposes by professionals only. It should not be used for clinical diagnosis or treatment, food or drug purposes, and should not be stored in residential areas.
• For your safety and health, please wear laboratory attire and disposable gloves while handling this product.

Storage

Store the product at -20°C, protected from light, to maintain its effectiveness for up to one year. For shorter-term storage, store at 4°C, also protected from light, for up to three months. If needed for immediate use, the product can be stored at room temperature for up to two weeks, still ensuring its quality.

Only for research and not intended for treatment of humans or animals

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