| Product Name | 30 min Western Assay Kit |
| Catalog No. | WBDK-YJL-0001 |
| 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 | 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 and secondary antibodies. 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 Procedure: After membrane transfer, immediately place the membrane (approx. 6.6×8.5 cm) into a western blot box containing 5 ml blocking Buffer. Shake gently at room temperature for 5 minutes. Note: For smaller membranes (~2×8.5 cm), use 2 ml blocking buffer and adjust reagent volumes proportionally. 2. Primary Antibody Incubation Preparation: Dilute the primary antibody according to the recommended dilution ratio. Procedure: Remove blocking buffer. Add 5 ml primary antibody working solution (2 ml for small membranes). Incubate at room temperature with gentle shaking for 15 minutes. Washing: Wash 3 times with 10 ml washing buffer, 30 seconds each (4 ml per wash for small membranes). Use a vacuum pump to remove excess buffer after each wash. 3. Secondary Antibody Incubation Preparation: Dilute secondary antibody at twice the recommended dilution ratio. Procedure: Add 5 ml secondary antibody working solution (2 ml for small membranes). Incubate at room temperature with gentle shaking for 5 minutes. Washing: Repeat 3 washes as described above. 4. Chemiluminescent Detection Preparation: Mix 0.5 ml chemiluminescent substrate reagent A and 0.5 ml chemiluminescent substrate reagent B to prepare 1 ml working solution (0.4 ml for small membranes). Use freshly prepared solution before detection. Procedure: Gently blot off excess liquid without touching the membrane surface. Place the membrane on cling film, add 1 ml working solution evenly, and incubate for 30 seconds. Detection: Remove excess solution, place membrane between two cling films, and proceed to chemiluminescence imaging or X-ray film exposure. 5. (Optional) Antibody Stripping and Housekeeping Protein Detection Stripping: Use neutral antibody stripping buffer to remove primary and secondary antibodies. Housekeeping Proteins: Use rapid detection kits to obtain target bands within 10 minutes. |
| Recommended Using Conditions | Using this product, high-quality western results of the target protein can usually be obtained by incubating at room temperature for 30 minutes, that is, blocking for 5 minutes, incubating with primary antibody for 15 minutes, washing 3 times for 30 seconds, incubating with secondary antibody for 5 minutes, washing 3 times for 30 seconds, incubating with the reagent for 30 seconds, and imaging is usually 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 (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 (much better than conventional blocking solution) |
| 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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