| Product Name | Superquick Western Assay Kit for Actin |
| Catalog No. | WBDK-YJL-0005 |
| Description | It only takes 10 minutes to get the internal reference protein Actin band from the transferred blot. |
| Detection Target | β-Actin |
| 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. Internal reference proteins are constitutively expressed in various tissues and cells, and their expression levels are relatively constant and rarely affected by external inducers. Therefore, they are widely used as a reference for whether the amount of Western protein loading is consistent. However, different cells, tissues, different conditions, and even different protein loading amounts will have a certain impact on the detection of internal reference proteins, so it is recommended to detect one or more internal reference proteins. |
| Packing List | Anti-Actin-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 the internal reference protein can usually be obtained within 10 minutes, which can greatly speed up the detection of the internal reference protein. |
| Application | It can be used for the detection of β-Actin in humans, mice, and rats. It is recommended to conduct preliminary experiments on other species. |
| Application Type | Western blot internal reference protein rapid detection |
| Quick Protocol | 1. Blocking Place the membrane (~6.6 × 8.5 cm) into a western box containing 5 ml blocking & antibody dilution buffer. Gently shake at room temperature for 1 minute. For smaller membrane (~2 × 8.5 cm), use 2 ml buffer. 2. Antibody Incubation Dilution: Prepare working solution by adding 10 μl Anti-Actin-HRP antibody into 5 ml buffer (1:500 dilution). Dilution can be adjusted from 1:250 to 1:1000 depending on signal strength. Incubation: After removing the blocking buffer, add 5 ml antibody solution and shake gently for 5 minutes at room temperature. Washing: Wash membrane 3 times, 30 seconds each, with 10 ml western wash buffer per wash. 3. Detection ECL Preparation: Mix 0.5 ml chemiluminescent substrate reagent A + 0.5 ml chemiluminescent substrate reagent B to prepare 1 ml working solution. For smaller membrane (~2 × 8.5 cm), use 0.4 ml. |
| 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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