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Cat. No.: RWGST-15 (for 6-15T)

Cat. No.: RWGST-75 (for 35-75T)

Description

Rapid Western Assay Kit for GST Tag is an ultra-fast Western blot detection kit that takes only 10 minutes to go from a completed transfer membrane to obtaining bands of GST-tagged fusion proteins. This kit includes Anti-GST Tag-HRP, Blocking and Antibody Dilution Buffer, Western wash buffer, and ECL Star chemiluminescent substrate. The primary and secondary antibody incubation is combined into a single step, making the process user-friendly with low background. It provides a complete set of reagents from blocking to chemiluminescence, enabling high-quality tag protein Western results in just 10 minutes post-transfer, greatly accelerating the detection of tagged proteins.

Tags like Flag, HA, Myc, His, GST, etc., are commonly used on expression vectors to facilitate the expression, purification, or detection of target proteins without the need for specific antibodies against the target protein. This series currently includes ultra-fast Western assay kits for Flag-tagged proteins, HA-tagged proteins, Myc-tagged proteins, His-tagged proteins, His-tagged proteins (C-terminal), GFP-tagged proteins, GST-tagged proteins, V5-tagged proteins, MBP-tagged proteins, StrepII-tagged proteins, and AVI-tagged proteins.

Blocking and Antibody Dilution Buffer in this kit can be used for both membrane blocking and antibody dilution. The blocking time is only 1 minute, with overall performance significantly superior to traditional blocking buffers based on BSA, non-fat milk, or casein. The diluted and prepared tag antibody working solution can be stored and used at 4ºC for at least one month and can be reused.

The kit uses Anti-GST Tag-HRP, so the incubation of primary and secondary antibodies is completed in one step. The antibody specificity is high, the sensitivity is excellent, and the optimized usage concentration is a 1:500 dilution. Typically, a 5-minute room temperature incubation is sufficient before proceeding to washing and detection.

The provided ECL Star chemiluminescent substrate in this kit offers high sensitivity, low background, and long-lasting luminescence, suitable for most conventional Western detections. If imaging needs optimization, higher sensitivity ECL Moon or ECL SuperMoon can be selected based on current experimental results.

Using this kit, the actual operation time from the completed transfer membrane is about 10 minutes.

Components

• Anti-GST Tag-HRP
• Blocking and Antibody Dilution Buffer
• Western Washing Buffer
• ECL Star Solution A
• ECL Star Solution B

Specification

If detecting a 6.6×8.5cm blot membrane, each Western blocking requires 5ml of Blocking and Antibody Dilution Buffer, and antibody dilution requires 5ml of Blocking and Antibody Dilution Buffer plus 10μl of antibody. The diluted antibody can be used a total of three times. Washing buffer is used at 10ml per wash, and the chemiluminescent reagent working solution requires 1ml per use. Based on this, the small and medium kit packages can perform 6 and 30 GST Tag Western rapid detections, respectively.

For the more common cut blot membranes of approximately 2×8.5cm, each blocking and antibody incubation requires 2ml of solution, 4ml of washing buffer per wash, and 0.4ml of chemiluminescent reagent working solution per use. The small and medium kit packages can perform 15 and 75 detections, respectively.

Storage Conditions

Store at -20ºC for one year. Blocking and Antibody Dilution Buffer, Western Washing Buffer, and ECL Star Solution A and B should be stored at 4ºC and are valid for at least six months.

Notes

• Please recycle the used antibody working solution. The recycled antibody working solution can be reused up to two more times. Store diluted antibodies, including used working solutions, at 4ºC.
• Recycled antibody working solutions should be used in the same way as freshly diluted antibodies. If slight turbidity is observed during reuse, centrifuge at 10,000×g for 1-3 minutes and use the supernatant for subsequent detection. If the recycled antibody working solution shows significant flocculation or mold/bacterial growth, consider discarding it.
• Using this product, high-quality internal reference protein Western results can typically be obtained within 10 minutes, which includes 1 minute for blocking, 5 minutes for antibody incubation, three 30-second washes, 1 minute for ECL reagent incubation, and imaging usually within 0.5 minutes.
• For 6.6×8.5cm blot membranes, it is recommended to use Western Washing Box for washing and incubation; for cut strip blots, 5-slot Western Washing Box is recommended.
• In some cases, to achieve better Western results, you may increase the time for blocking, antibody incubation, and washing, and optimize the exposure time for imaging as needed.
• Both ECL Star Solution A and B are harmful to humans. Handle with care and ensure effective protection to avoid direct contact with skin or inhalation.
• This product is for scientific research use by professionals only. It is not intended for clinical diagnosis or treatment, food, or drug use, and should not be stored in a residential setting.
• For your safety and health, please wear a lab coat and disposable gloves during operation.