| Product Name | 30 min Western Assay Kit for Mouse Primary Antibody |
| Catalog No. | WBDK-YJL-0003 |
| Description | It usually takes only 30 minutes to obtain the target protein band from the transferred blotting membrane. |
| Detection Method | Chemiluminescence |
| Storage | Store at -20ºC for up to one year, or at 4ºC for at least three months. |
| 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. |
| Packing List | HRP-labeled goat anti-mouse IgG (H+L), Blocking solution, antibody diluent, western washing solution, chemiluminescent substrate reagent A solution, chemiluminescent substrate reagent B solution |
| Features | This kit is easy to use and can significantly shorten the time for blocking, primary antibody and secondary antibody incubation. The kit contains all the solutions required from blocking to chemiluminescence. Users only need to prepare their own primary antibody. After the transfer is completed, high-quality Western results of the target protein can usually be obtained within 30 minutes, which can greatly speed up the detection of the target protein. |
| Application | Ideal for routine and high-throughput western blotting requiring fast and efficient target protein detection. |
| Required Materials And Equipment (Not Included) | Primary antibody, secondary antibody, water, anhydrous ethanol, and conventional equipment such as electrophoresis, transfer and imaging |
| Quick Protocol | 1. Membrane blocking Place the membrane (6.6×8.5 cm) into 5 ml blocking buffer in a tray, gently shake at room temperature for 5 minutes. For sliced membranes (~2×8.5 cm), use 2 ml. 2. Primary Antibody Incubation Prepare the working solution. Remove blocking buffer and add 5 ml primary antibody solution, incubate at room temp for 15 min, or at 4ºC for 15–30 min. Recover antibody, wash membrane 3× with 10 ml wash buffer, 30 sec each. For small strips, use 2 ml antibody and 4 ml wash buffer per wash. 3. Secondary Antibody Incubation Prepare 2° antibody: 5 ml diluent + 10 µl HRP-labeled goat anti-mouse IgG(H+L). Incubate 5 ml solution at room temp for 5 min, or at 4ºC for 5–10 min. Recover antibody, wash membrane 3× with 10 ml wash buffer, 30 sec each. 4. Chemiluminescent Detection Mix 0.5 ml ECL A + 0.5 ml ECL B to prepare 1 ml working solution right before use. Cover membrane with 1 ml ECL, react for 30 sec at room temperature. Blot excess liquid and proceed to imaging. 5. (Optional) Stripping & Internal Control Detection Use stripping buffer to remove antibodies. For internal control detection. |
| Recommended Using Conditions | Blocking for 5 minutes, primary antibody incubation for 15 minutes, washing for 30 seconds × 3 times, secondary antibody incubation for 5 minutes, washing for 30 seconds × 3 times, incubation with the reagent for 30 seconds, and imaging typically within 0.5 minutes. |
| Hazard Statements | Chemiluminescent substrate reagents A and B are harmful to the human body and require protection during operation. |
| Total Time of Experiment | About 30 minutes |
| Compatible Temperature Conditions | Primary/secondary antibodies can be incubated at 4ºC: primary antibody 15–30 minutes, secondary antibody 5–10 minutes. |
| Blocking Solution Action Time | 5 minutes |
| Blocking Solution Features | The overall effect is significantly better than traditional blocking solutions based on BSA (bovine serum albumin), skimmed milk powder, casein, etc. |
| Antibody Diluent Features | Suitable for both primary and secondary antibodies, can be reused at least 2 times, and can be stored at 4ºC for ≥1 month |
| Washing Solution Application | Used for washing after incubation of primary or secondary antibodies at Western time |
| Washing Solution Action Time | 30 seconds per wash can reduce background and improve signal-to-noise ratio |
| 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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