| Product Name | 30 min Western Assay Kit for Rabbit Primary Antibody |
| Catalog No. | WBDK-YJL-0004 |
| 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-rabbit 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. Blocking After transfer, place the membrane (~6.6 × 8.5 cm) into a western box containing 5 ml blocking buffer. Shake gently at room temperature for 5 minutes. For smaller membrane (~2 × 8.5 cm), use 2 ml buffer. 2. Primary Antibody Incubation Dilution: Dilute primary antibody in 5 ml antibody diluent according to recommended ratio. Incubation: After removing blocking buffer, add 5 ml antibody solution and incubate at room temperature on a shaker for 15 minutes. Washing: Wash the membrane 3 times, 30 seconds each, with 10 ml wash buffer per wash. Use vacuum to remove excess liquid. 3. Secondary Antibody Incubation Dilution: Prepare 5 ml of working solution by mixing 10 μl HRP-conjugated goat anti-Rabbit IgG (H+L) with 5 ml antibody diluent. Incubation: Shake at room temperature for 5 minutes. Washing: Wash 3 times, 30 seconds each, with 10 ml wash buffer. 4. Detection Prepare ECL Working Solution: Mix 0.5 ml Solution A + 0.5 ml Solution B to make 1 ml chemiluminescent substrate reagent reagent. Apply: Use tweezers to place membrane on plastic wrap. Add 1 ml chemiluminescent substrate reagent solution and incubate for 30 seconds. Image: Blot excess liquid, wrap membrane, and proceed with chemiluminescent imaging. 5. (Optional) Antibody Stripping & Loading Control Detection To reuse the membrane for detecting other targets, use neutral pH Antibody Stripping Buffer. For fast detection of loading controls, use 10-minute Quick Western Kits. |
| Recommended Using Conditions | Blocking for 5 min, primary antibody incubation for 15 min, wash 3×30 s, secondary antibody incubation for 5 min, wash 3×30 s, incubation for 30 s, 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 30 minutes (from transfer to obtaining the target protein band) |
| 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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