| Product Name | Superquick Western Assay Kit for AVI Tag |
| Catalog No. | WBDK-YJL-0018 |
| Description | It takes only 10 minutes from the transfer of the blot to the acquisition of AVI-tagged fusion protein bands. |
| Detection Target | AVI tag fusion protein |
| Detection Method | Chemiluminescence |
| Storage | Store at -20ºC for up to one year. |
| Intended Use | For research use only. |
| Results | Clear bands, low background, high sensitivity |
| Background | Western, also known as western blot, western blotting, often abbreviated as WB, is one of the important methods for detecting protein levels using antibodies. Flag tag, HA tag, Myc tag, His tag, GST tag, etc. are some of the most common tags on expression vectors. Fusion expression with these tags can be very conveniently used for the expression, purification or detection of target proteins without the need for target protein-specific antibodies. |
| Packing List | Anti-AVI Tag-HRP, blocking and antibody diluent, western washing solution, chemiluminescent substrate reagent A solution, chemiluminescent substrate reagent B solution |
| Features | The primary antibody and secondary antibody incubation are combined into a one-step operation, which is easy to use and has low background. It includes a complete set of reagents from blocking to chemiluminescence. After the transfer is completed, high-quality western results of tagged proteins can usually be obtained within 10 minutes, which can greatly speed up the detection of tagged proteins. |
| Application Type | Western blot rapid detection of tagged proteins |
| Quick Protocol | 1. Membrane blocking After transfer, immediately place the membrane (~6.6×8.5 cm) into a western washing box containing 5 ml blocking and antibody dilution buffer. Place on a shaker and gently shake at room temperature for 1 minute. For smaller membranes (~2×8.5 cm), use 2 ml blocking buffer. Note: 1-minute blocking at room temperature is usually sufficient. For more blocking buffer, use western blocking Buffer. 2. Antibody Incubation Prepare Anti-AVI Tag-HRP antibody working solution by diluting 1:500 (e.g., add 10 μl antibody to 5 ml buffer). Adjust dilution between 1:250 and 1:1000 if needed based on protein amount. More antibody dilution buffer available. Remove blocking buffer using a vacuum pump, pipette, or dropper. Add 5 ml antibody working solution (2 ml for smaller membranes). Incubate on shaker at room temperature for 5 minutes. Save antibody solution for reuse. Wash membrane 3 times with Western wash buffer, 10 ml each time for 30 seconds (4 ml for smaller membranes). More wash buffer available. 3. Protein Detection Prepare chemiluminescent substrate reagent working solution by mixing 0.5 ml chemiluminescent substrate reagent A and 0.5 ml chemiluminescent substrate reagent B just before use (0.4 ml for smaller membranes). Remove membrane with WB tweezers, gently blot excess liquid without touching protein side, place on clean plastic wrap. Add 1 ml chemiluminescent substrate reagent solution evenly on membrane (0.4 ml for smaller membranes), incubate for 1 minute. Remove solution, blot excess liquid, sandwich membrane between two plastic wraps. Proceed with chemiluminescence imaging or film exposure. |
| Recommended Using Conditions | Blocking for 1 min, antibody incubation for 5 min, wash 3×30 s, incubation for 1 min, imaging within 0.5 min. |
| Hazard Statements | Chemiluminescent substrate reagents A and B are harmful to the human body and require protection during operation. |
| Total Time of Experiment | About 10 minutes (from transfer to obtaining the target protein band) |
| Compatible Temperature Conditions | Primary and secondary antibody incubations are combined into a single step, usually incubated for 5 minutes at room temperature before subsequent washing and detection. |
| Blocking Solution Action Time | 1 minute |
| Blocking and Antibody Dilution Buffer Features | Can be used for both membrane blocking and antibody dilution simultaneously. |
| Developer Features | High sensitivity, low background, long-term luminescence |
| Recommended Supporting Consumables | Western membrane washing box: 9.0×6.0×3.3 cm; 5 grids, 14.5×9.8×3.5 cm |
| Blotting Membrane Sizes and Recommended Usage Times | 6.6×8.5 cm blotting membrane: 6 times 2×8.5 cm blotting membrane: 15 times |
| Note | For your safety and health, please wear a lab coat and disposable gloves during the operation. |
For research use only, not for clinical use.
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