| Product Name | Superquick Western Assay Kit for Flag Tag |
| Catalog No. | WBDK-YJL-0008 |
| Description | It only takes 10 minutes to get the fusion protein band with Flag tag from the transferred blot. |
| Detection Target | Flag 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-Flag 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 Place the membrane in a container with 5 ml blocking & antibody dilution buffer. Gently rock at room temperature for 1 minute. Use 2 ml buffer for ~2 × 8.5 cm cut membranes. Note: blocking time can be extended as needed. 2. Antibody Incubation Prepare antibody solution: Add 10 μl Anti-Flag Tag-HRP to 5 ml buffer (1:500 dilution). Dilution range: 1:250 to 1:1000 depending on protein expression. Remove blocking buffer completely. Add 5 ml antibody solution (or 2 ml for smaller membranes). Incubate with gentle rocking at room temperature for 5 minutes. Recover antibody solution for reuse. Wash the membrane 3 times, 30 seconds each with 10 ml western wash buffer (or 4 ml for smaller membranes). 3. Detection Prepare ECL working solution: mix 0.5 ml reagent A + 0.5 ml reagent B. Use 0.4 ml total for small membranes. Remove excess liquid from the membrane and place on clean plastic wrap. Add 1 ml ECL working solution, ensure even coverage, and let sit for 1 minute. Discard excess solution, gently blot dry, seal in wrap, and image with chemiluminescent imager or film. |
| 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 or 30 times 2×8.5 cm blotting membrane: 15 times or 75 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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