| Product Name | Superquick Western Assay Kit for V5 Tag |
| Catalog No. | WBDK-YJL-0015 |
| Description | It only takes 10 minutes to get from the blot to the V5-tagged fusion protein band. |
| Detection Target | V5 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-V5 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. Blocking After transfer, place the membrane into a western washing box containing 5 ml blocking and antibody dilution buffer. Shake gently at room temperature on a shaker for 1 minute. For smaller membranes (~2×8.5 cm), use 2 ml blocking buffer. Note: 1 minute blocking is usually sufficient. Use more blocking solution if needed. 2. Antibody Incubation Prepare antibody working solution by diluting Anti-V5 Tag-HRP antibody 1:500 (e.g., add 10 µl antibody to 5 ml blocking buffer). Dilution range: 1:250 to 1:1000 can be adjusted based on protein amount. More dilution buffer available. Remove blocking buffer using vacuum pump or pipette. Add 5 ml antibody working solution (2 ml for smaller membranes). Incubate at room temperature with gentle shaking 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 each of solution A and B, room temperature before use (0.4 ml for smaller membranes). Remove membrane with tweezers, gently blot excess liquid without touching the protein side, and place on clean plastic wrap. Add 1 ml chemiluminescent substrate reagent working solution evenly on membrane (0.4 ml for smaller membranes), incubate 1 minute. Remove solution, blot excess, place membrane between two plastic wraps. Proceed to chemiluminescence imaging or film detection. |
| 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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